Nitroso compounds as nitroxyl donors and methods of use thereof

ABSTRACT

The invention relates to nitroso derivatives including carboxylic acid and phosphoric acid esters of hydroxy nitroso compounds that donate nitroxyl (HNO) under physiological conditions. The compounds and compositions of the invention are useful in treating and/or preventing the onset and/or development of diseases or conditions that are responsive to nitroxyl therapy, including heart failure, ischemia/reperfusion injury and cancer.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Patent ApplicationNo. 61/051,287 filed May 7, 2008, the disclosures of which isincorporated herein by reference in its entirety.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

Not applicable.

BACKGROUND OF THE INVENTION

Heart Failure

Congestive heart failure (CHF) is a generally progressive, lifethreatening condition in which myocardial contractility is depressedsuch that the heart is unable to adequately pump the blood returning toit, also referred to as decompensation. Symptoms include breathlessness,fatigue, weakness, leg swelling, and exercise intolerance. On physicalexamination, patients with heart failure often have elevated heart andrespiratory rates (an indication of fluid in the lungs), edema, jugularvenous distension, and/or enlarged hearts. The most common cause of CHFis atherosclerosis, which causes blockages in the coronary arteries thatprovide blood flow to the heart muscle. Ultimately, such blockages maycause myocardial infarction with subsequent decline in heart functionand resultant heart failure. Other causes of CHF include valvular heartdisease, hypertension, viral infections of the heart, alcoholconsumption, and diabetes. Some cases of CHF occur without clearetiology and are called idiopathic. The effects of CHF on a subjectexperiencing the condition can be fatal.

There are several types of CHF. Two types of CHF are identifiedaccording to which phase of the cardiac pumping cycle is more affected.Systolic heart failure occurs when the heart's ability to contractdecreases. The heart cannot pump with enough force to push a sufficientamount of blood into the circulation leading to a reduced leftventricular ejection fraction. Lung congestion is a typical symptom ofsystolic heart failure. Diastolic heart failure refers to the heart'sinability to relax between contractions and allow enough blood to enterthe ventricles. Higher filling pressures are required to maintaincardiac output, but contractility as measured by left ventricularejection fraction is typically normal. Swelling (edema) in the abdomenand legs is a typical symptom of diastolic heart failure. Often, anindividual experiencing heart failure will have some degree of bothsystolic heart failure and diastolic heart failure.

CHF is also classified according to its severity. The New York HeartAssociation classifies CHF into four classes: Class I involves noobvious symptoms, with no limitations on physical activity; Class IIinvolves some symptoms during or after normal activity, with mildphysical activity limitations; Class III involves symptoms with lessthan ordinary activity, with moderate to significant physical activitylimitations; and Class IV involves significant symptoms at rest, withsevere to total physical activity limitations. Typically, an individualprogresses through the classes as they live with the condition.

Although CHF is generally thought of as a chronic, progressivecondition, it can also develop suddenly. This type of CHF is calledacute CHF, and it is a medical emergency. Acute CHF can be caused byacute myocardial injury that affects either myocardial performance, suchas myocardial infarction, or valvular/chamber integrity, such as mitralregurgitation or ventricular septal rupture, which leads to an acuterise in left ventricular and diastolic pressure resulting in pulmonaryedema and dyspnea.

Common treatment agents for CHF include vasodilators (drugs that dilateblood vessels), positive inotropes (drugs that increase the heart'sability to contract), and diuretics (drugs to reduce fluid).Additionally, beta-antagonists (drugs that antagonize beta-adrenergicreceptors) have become standard agents for treating mild to moderateheart failure. Lowes et al. Clin. Cardiol., 23:III11-6 (2000).

Positive inotropic agents include beta-adrenergic agonists, such asdopamine, dobutamine, dopexamine, and isoproterenol. However, use of abeta-agonist has potential complications, such as arrhythmogenesis andincreased oxygen demand by the heart. Additionally, the initialshort-lived improvement of myocardial contractility afforded by thesedrugs is followed by an accelerated mortality rate resulting largelyfrom a greater frequency of sudden death. Katz, HEART FAILURE:PATHOPHYSIOLOGY, MOLECULAR BIOLOGY AND CLINICAL MANAGEMENT, Lippincott,Williams & Wilkins (1999).

Beta-antagonists antagonize beta-adrenergic receptor function. Whileinitially contra-indicated in heart failure, they have been found toprovide a marked reduction in mortality and morbidity in clinicaltrials. Bouzamondo et al., Fundam. Clin. Pharmacol., 15: 95-109 (2001).Accordingly, they have become an established therapy for heart failure.However, even subjects that improve under beta-antagonist therapy maysubsequently decompensate and require acute treatment with a positiveinotropic agent. Unfortunately, as their name suggests, beta-antagonistsblock the mechanism of action of the positive inotropic beta-agoniststhat are used in emergency care centers. Bristow et al., J. Card. Fail.,7: 8-12 (2001).

Vasodilators, such as nitroglycerin, have been used for a long period oftime to treat heart failure. However, the cause of nitroglycerin'stherapeutic effect was not known until late in the last century when itwas discovered that the nitric oxide molecule (NO) was responsible fornitroglycerin's beneficial effects. In some subjects experiencing heartfailure, a nitric oxide donor is administered in combination with apositive inotropic agent to both cause vasodilation and to increasemyocardial contractility. However, this combined administration canimpair the effectiveness of positive inotropic treatment agents. Forexample, Hart et al, Am. J. Physiol. Heart Circ. Physiol., 281:146-54(2001) reported that administration of the nitric oxide donor sodiumnitroprusside, in combination with the positive inotropic,beta-adrenergic agonist dobutamine, impaired the positive inotropiceffect of dobutamine. Hare et al., Circulation, 92:2198-203 (1995) alsodisclosed the inhibitory effect of nitric oxide on the effectiveness ofdobutamine.

As described in U.S. Pat. No. 6,936,639, compounds that donate nitroxyl(HNO) under physiological conditions have both positive inotropic andlusotropic effects and offer significant advantages over existingtreatments for failing hearts. Due to their concomitant positiveinotropic/lusotropic action and unloading effects, nitroxyl donors werereported as helpful in treating cardiovascular diseases characterized byhigh resistive load and poor contractile performance. In particular,nitroxyl-donating compounds were reported as useful in the treatment ofheart failure, including heart failure in individuals receivingbeta-antagonist therapy.

Ischemia

Ischemia is a condition characterized by an interruption or inadequatesupply of blood to tissue, which causes oxygen deprivation in theaffected tissue. Myocardial ischemia is a condition caused by a blockageor constriction of one or more of the coronary arteries, such as canoccur with atherosclerotic plaque occlusion or rupture. The blockade orconstriction causes oxygen deprivation of the non-perfused tissue, whichcan cause tissue damage. Further, upon reperfusion with subsequentreoxygenation of the tissue, when the blood is able to flow again or theoxygen demand of the tissue subsides, additional injury can be caused byoxidative stress.

Ischemia/reperfusion injury refers to tissue damage caused by oxygendeprivation followed by reoxygenation. The effects ofischemia/reperfusion injury in a subject experiencing the condition canbe fatal, particularly when the injury occurs in a critical organ suchas the heart or brain.

Accordingly, compounds and compositions effective in preventing orprotecting against ischemia/reperfusion injury would be usefulpharmaceuticals. Compounds such as nitroglycerin have been used for along period of time to help control vascular tone and protect againstmyocardial ischemia/reperfusion injury. It was discovered that thenitric oxide molecule was responsible for nitroglycerin's beneficialeffects. This discovery prompted interest in medical uses for nitricoxide and investigations into related species such as nitroxyl. Asreported in U.S. patent application Ser. No. 10/463,084 (U.S.Publication No. 2004/0038947) administration of a compound that donatesnitroxyl under physiological conditions, prior to ischemia, canattenuate ischemia/reperfusion injury to tissues, for example,myocardial tissues. This beneficial effect was reported as a surprisingresult given that nitroxyl was previously reported to increaseischemia/reperfusion injury (See, Ma et al., “Opposite Effects of NitricOxide and Nitroxyl on Postischemic Myocardial Injury,” Proc. Nat'l Acad.Sci., 96(25): 14617-14622 (1999), reporting that administration ofAngeli's salt (a nitroxyl donor under physiological conditions) toanesthetized rabbits during ischemia and 5 minutes prior to reperfusionincreased myocardial ischemia/reperfusion injury and Takahira et al.,“Dexamethasone Attenuates Neutrophil Infiltration in the Rat Kidney inIschemia/Reperfusion Injury: The Possible Role of Nitroxyl,” FreeRadical Biology & Medicine, 31(6):809-815 (2001) reporting thatadministration of Angeli's salt during ischemia and 5 minutes beforereperfusion of rat renal tissue contributed to neutrophil infiltrationinto the tissue, which is believed to mediate ischemia/reperfusioninjury). In particular, pre-ischemic administration of Angeli's salt andisopropylamine/NO has been reported to prevent or reduceischemia/reperfusion injury.

Cancer

One of the challenges in developing anti-cancer drugs is to discovercompounds that are selectively toxic to tumor cells over normal cells.It has been found that tumor tissues have an acidic microenvironmentwith a pH from 6.0 to 7.0, while the extra- and intracellular milieu ofnormal cells has a pH of 7.4. Angeli's salt has been reported to exhibitstrong cytotoxicity to cancer cells in weakly acidic solutions, whereasno toxicity was observed at pH 7.4 (Stoyanovsky, D. A. et al. J. Med.Chem. 2004, 47, 210-217; and WO 03/020221). In a subcutaneous xenograftmodel of pheochromocytoma, Angeli's salt was found to inhibit tumorgrowth at a dose that was nontoxic to nude mice. Nitroxyl derivativesthat are not known to release HNO, such as ruboxyl, a nitroxyl analogueof daunorubicin, have been shown to be active against hepatic metastasesfrom colorectal carcinoma (Sirovich, I. et al Tumor Biol. 1999;20:270-276).

Norris A. J. et al (Intl. J. Cancer 2008, 122, 1905-1910) reported thatAngeli's salt inhibits the proliferation of cultured breast cancer cellsand decreases tumor mass in a mouse xenograft model. Norris A. J. et alproposed that HNO released from Angeli's salt blocks glycolysis incancer cells by inhibiting the enzyme glyceraldehyde 3-phosphatedehydrogenase (GAPDH), resulting in decreased levels of HIF-1α(hypoxia-inducible factor) protein and activity, lower VEGF (vascularendothelial growth factor) production, decreased tumor angiogenesis andan increase in apoptotic cells.

Nitroxyl Donors

Due to its inherent reactivity, HNO must be generated in situ from donorcompounds. To date, the vast majority of studies of the biologicaleffect of HNO have used the donor sodium α-oxyhyponitrite (“Angeli'ssalt” or “AS”). However, the chemical stability of AS has made itunsuitable to develop as a therapeutic agent. Angeli's salt alsoreleases nitrite, which possesses its own biological profile.N-hydroxybenzenesulfonamide (“Piloty's acid” or “PA”) has previouslybeen shown to be a nitroxyl donor only at high pH (>9) (Bonner, F. T.;Ko, Y. Inorg. Chem. 1992, 31, 2514-2519). Under physiologicalconditions, PA has been shown to be a nitric oxide donor via anoxidative pathway (Zamora, R.; Grzesiok, A.; Weber, H.; Feelisch, M.Biochem. J. 1995, 312, 333-339). International Patent ApplicationPublication WO 2007/109175 describes N-hydroxylsulfonamide derivativesthat donate nitroxyl under physiological conditions.

Acyloxy nitroso compounds have been reported to yield nitroxyl in situwhen reacted with nucleophiles (Sha, X. et al J. Am. Chem. Soc. 2006,128, 9687-9692). Although Rehse and Herpel (Arch. Pharm. Med. Chem.1998, 331, 104-110) showed acyloxy nitroso compounds inhibit plateletaggregation and thrombus formation (indicative of NO release), theygenerate only small amounts (<1%) of NO and HNO under neutralconditions. International Patent Application Publication WO 2007/120839describes conjugates of acyloxy nitroso compounds with non-steroidalanti-inflammatory drugs (NSAID) as nitroxyl donors for treatingcongestive heart failure.

Significant Medical Need

Despite efforts towards the development of new therapies for thetreatment of diseases and conditions such as heart failure,ischemia/reperfusion injury and cancer, there remains a significantinterest in and need for additional or alternative compounds that treator prevent the onset or severity of these and related diseases orconditions. In particular, there remains a significant medical need foralternative or additional therapies for the treatment of diseases orconditions that are responsive to nitroxyl therapy. New compounds thatdonate nitroxyl under physiological conditions and methods of usingcompounds that donate nitroxyl under physiological conditions may thusfind use as therapies for treating, preventing and/or delaying the onsetand/or development of diseases or conditions responsive to nitroxyltherapy, including heart disease, ischemia/reperfusion injury andcancer. Preferably, the therapeutic agents can improve the quality oflife and/or prolong the survival time for patients with the disease orcondition.

BRIEF SUMMARY OF THE INVENTION

The invention provides nitroxyl donor compounds and compositions thatare useful in treating and/or preventing the onset and/or development ofdiseases or conditions that are responsive to nitroxyl therapy, such asheart failure, ischemia/reperfusion injury and cancer.

In one aspect, the invention provides compounds of the formula (I):

where each R¹ and R² is independently a substituted or unsubstitutedC₁-C₈ alkyl, substituted or unsubstituted C₂-C₈ alkenyl, substituted orunsubstituted C₂-C₈ alkynyl, or R¹ and R² are taken together to form anunsubstituted or substituted 6 or 7-membered carbocyclic moiety or anunsubstituted or substituted 5, 6 or 7-membered heterocyclic moiety; andD is selected from the group consisting of alkyl-C(O)—, substitutedalkyl-C(O)—, perhaloalkyl-C(O)—, alkenyl-C(O)—, substitutedalkenyl-C(O)—, alkynyl-C(O)—, substituted alkynyl-C(O)—, aryl-C(O)—,substituted aryl-C(O)—, heteroaryl-C(O)—, substituted heteroaryl-C(O)—,heterocyclyl-C(O)— and —P(O)(OC₁-C₈ alkyl)₂; provided that the compoundis other than 1-nitrosocycloheptyl acetate, 1-nitrosocycloheptylbenzoate, 9-nitrosobicyclo[3.3.1]nonan-9-yl acetate or8-methyl-3-nitroso-8-azabicyclo[3.2.1]octan-3-yl acetate; and (i) whenR¹ or R² is an unsubstituted C₁-C₈ alkyl, the R¹ or R² that is anunsubstituted C₁-C₈ alkyl is other than methyl or propyl and D is otherthan an NSAID moiety; (ii) when R¹ and R² are taken together to form anunsubstituted 6-membered carbocyclic moiety, D is other thann-alkyl-C(O)—, ClCH₂—C(O)—, CCl₃—C(O)—, CF₃—C(O)—, (CH₃)₃C—C(O)—,alkenyl-C(O)—, substituted alkenyl-C(O)—, unsubstituted aryl-C(O)—,mono-substituted aryl-C(O)— or an NSAID moiety; (iii) when R¹ and R² aretaken together to form a substituted 6-membered carbocyclic moiety, thesubstituted 6-membered carbocyclic moiety is a monocyclic or bicyclicring that is substituted with a moiety other than alkyl, nitroso, acyl,oxime, and substituted alkenyl; or (iv) when R¹ and R² are takentogether to form an unsubstituted or substituted 5 or 6-memberedheterocyclic moiety, the 5 or 6-membered heterocyclic moiety is amonocyclic or bicyclic ring other than dioxane or acyloxy-substitutedtetrahydropyan, or salts or solvates thereof.

In one variation, the invention provides a method of treating a diseaseor condition that is responsive to nitroxyl therapy, such as treating anindividual who has heart failure, ischemia/reperfusion injury or cancerby administering to the individual a therapeutically effective amount ofa compound of formula (I), where each R¹ and R² is independently asubstituted or unsubstituted C₁-C₈ alkyl, substituted or unsubstitutedC₂-C₈ alkenyl, substituted or unsubstituted C₂-C₈ alkynyl, or R¹ and R²are taken together to form an unsubstituted or substituted 6 or7-membered carbocyclic moiety or an unsubstituted or substituted 5, 6 or7-membered heterocyclic moiety; and D is selected from the groupconsisting of alkyl-C(O)—, substituted alkyl-C(O)—, perhaloalkyl-C(O)—,alkenyl-C(O)—, substituted alkenyl-C(O)—, alkynyl-C(O)—, substitutedalkynyl-C(O)—, aryl-C(O)—, substituted aryl-C(O)—, heteroaryl-C(O)—,substituted heteroaryl-C(O)—, heterocyclyl-C(O)— and—P(O)(OC₁-C₈alkyl)₂.

In one embodiment, the invention embraces compounds of the formula (II):

where D is selected from the group consisting of alkyl-C(O)—,substituted alkyl-C(O)—, perhaloalkyl-C(O)—, alkenyl-C(O)—, substitutedalkenyl-C(O)—, alkynyl-C(O)—, substituted alkynyl-C(O)—, aryl-C(O)—,substituted aryl-C(O)—, heteroaryl-C(O)—, substituted heteroaryl-C(O)—,heterocyclyl-C(O)—and —P(O)(OC₁-C₈alkyl)₂; X is O, NR⁴, CR⁵R⁶, S, S(O)or S(O)₂; Y is CR⁵R⁶ or CR⁵R⁶—CR⁷R⁸; Z is CR⁵R⁶ or a bond, provided thatwhen X is CR⁵R⁶, Z is CR⁵R⁶; R⁴ is H, substituted or unsubstituted C₁-C₈alkyl, substituted or unsubstituted C₂-C₈ alkenyl, substituted orunsubstituted C₂-C₈ alkynyl, substituted or unsubstituted acyl,alkoxycarbonyl, substituted or unsubstituted heterocyclyl, substitutedor unsubstituted aryl, substituted or unsubstituted aralkyl, orsulfonyl; and each R⁵, R⁶, R⁷, R⁸, R^(11a), R^(11b), R^(12a) and R^(12b)is independently H, substituted or unsubstituted C₁-C₈ alkyl, halo,hydroxyl, alkoxy, cyano, nitro, or is taken together with a geminal Rgroup to form a carbonyl moiety, or is taken together with a vicinal Rgroup to form a bond, or is taken together with another R group to forma ring; provided that the compound is other than 1-nitrosocycloheptylacetate, 1-nitrosocycloheptyl benzoate,9-nitrosobicyclo[3.3.1]nonan-9-yl acetate or8-methyl-3-nitroso-8-azabicyclo[3.2.1]octan-3-yl acetate; and (i) whenX, Y, Z, R^(11a), R^(11b), R^(12a), and R^(12b) are taken together toform an unsubstituted 6-membered carbocyclic moiety, D is other thann-alkyl-C(O)—, ClCH₂—C(O)—, CCl₃—C(O)—, CF₃—C(O)—, (CH₃)₃C—C(O)—,alkenyl-C(O)—, substituted alkenyl-C(O)—, unsubstituted aryl-C(O)—,mono-substituted aryl-C(O)— or an NSAID moiety; (ii) when X, Y, Z,R^(11a), R^(11b), R^(12a), and R^(12b) are taken together to form asubstituted 6-membered carbocyclic moiety, the substituted 6-memberedcarbocyclic moiety is a monocyclic or bicyclic ring that is substitutedwith a moiety other than alkyl, nitroso, acyl, oxime, and substitutedalkenyl; or (iii) when X, Y, Z, R^(11a), R^(11b), R^(12a), and R^(12b)are taken together to form an unsubstituted or substituted 5 or6-membered heterocyclic moiety, the 5 or 6-membered heterocyclic moietyis a monocyclic or bicyclic ring other than dioxane oracyloxy-substituted tetrahydropyran; or salts or solvates thereof. Inone variation, the compound is of formula (II) where X is O, NR⁴, S,S(O) or S(O)₂.

In one variation, the invention provides a method of treating a diseaseor condition that is responsive to nitroxyl therapy, such as treating anindividual who has heart failure, ischemia/reperfusion injury or cancerby administering to the individual a therapeutically effective amount ofa compound of formula (II), where D is selected from the groupconsisting of alkyl-C(O)—, substituted alkyl-C(O)—, perhaloalkyl-C(O)—,alkenyl-C(O)—, substituted alkenyl-C(O)—, alkynyl-C(O)—, substitutedalkynyl-C(O)—, aryl-C(O)—, substituted aryl-C(O)—, heteroaryl-C(O)—,substituted heteroaryl-C(O)—, heterocyclyl-C(O)— and—P(O)(OC₁-C₈alkyl)₂; X is O, NR⁴, CR⁵R⁶, S, S(O) or S(O)₂; Y is CR⁵R⁶ orCR⁵R⁶—CR⁷R⁸; Z is CR⁵R⁶ or a bond, provided that when X is CR⁵R⁶, Z isCR⁵R⁶; R⁴ is H, substituted or unsubstituted C₁-C₈ alkyl, substituted orunsubstituted C₂-C₈ alkenyl, substituted or unsubstituted C₂-C₈ alkynyl,substituted or unsubstituted acyl, alkoxycarbonyl, substituted orunsubstituted heterocyclyl, substituted or unsubstituted aryl,substituted or unsubstituted aralkyl, or sulfonyl; and each R⁵, R⁶, R⁷,R⁸, R^(11a), R^(11b), R^(12a) and R^(12b) is independently H,substituted or unsubstituted C₁-C₈ alkyl, halo, hydroxyl, alkoxy, cyano,nitro, or is taken together with a geminal R group to form a carbonylmoiety, or is taken together with a vicinal R group to form a bond, oris taken together with another R group to form a ring.

In another aspect, the invention provides a pharmaceutical compositioncomprising a compound of the invention and a pharmaceutically acceptablecarrier. In one embodiment, the pharmaceutical composition is suitablefor parenteral administration, e.g., aqueous or non-aqueous sterileinjection solutions. In one variation, the pharmaceutical composition isacidic, e.g. an aqueous composition having a pH of about 5.5 to about 7.

In yet another aspect, the invention provides a method of treating adisease or condition that is responsive to nitroxyl therapy comprisingadministering to an individual having a disease or condition that isresponsive to nitroxyl therapy a therapeutically effective amount of acompound of the invention or a pharmaceutically acceptable salt thereof.This invention embraces methods of delivering a therapeuticallyeffective amount of nitroxyl administering a compound of the inventionin a suitable dose. In one embodiment, the condition is heart failure.In another embodiment, the condition is ischemia/reperfusion injury. Inone embodiment, the invention provides a method for treating a patienthaving a cancerous disease comprising administering to the patient atherapeutically effective amount of a compound of the invention or apharmaceutically acceptable salt thereof. In another embodiment, thecancerous disease is a breast cancer, a pancreatic cancer, a prostatecancer or a colorectal cancer.

DETAILED DESCRIPTION OF THE INVENTION Definitions

Unless clearly indicated otherwise, the following terms as used hereinhave the meanings indicated below.

Use of the terms “a”, “an” and the like refers to one or more.

“Alkyl” intends a linear saturated hydrocarbon structure having 1 to 20carbon atoms, preferably 1 to 12 carbon atoms and more preferably 1 to 8carbon atoms or 1 to 4 carbon atoms. “Alkyl” also intends a branched orcyclic hydrocarbon structure having 3 to 20 carbon atoms, preferably 3to 12 carbon atoms and more preferably 3 to 8 carbon atoms. For any useof the term “alkyl,” unless clearly indicated otherwise, it is intendedto embrace all variations of alkyl groups disclosed herein, as measuredby the number of carbon atoms, the same as if each and every alkyl groupwas explicitly and individually listed for each usage of the term. Whenthe alkyl group is cyclic, it may also be referred to as a cycloalkylgroup and have e.g., 3 to 20 annular carbon atoms, preferably 3 to 12annular carbon atoms and more preferably 3 to 8 annular carbon atoms.When an alkyl residue having a specific number of carbons is named, allgeometric isomers having that number of carbons are intended to beencompassed; thus, for example, “butyl” is meant to include n-butyl,sec-butyl, iso-butyl and t-butyl; “propyl” includes n-propyl andiso-propyl. Examples of alkyl groups include methyl, ethyl, n-propyl,i-propyl, t-butyl, n-heptyl, octyl, cyclopentyl, cyclopropyl,cyclobutyl, norbornyl, and the like. Alkyl is also used herein to denotean alkyl residue as part of a larger functional group and when so used,is taken together with other atoms to form another functional group. Forinstance, reference to —C(O)O-alkyl intends an ester functional group,where the alkyl portion of the moiety may be any alkyl group, andprovide by way of example only, the functional group —C(O)OCH₃,—C(O)OCH(CH₃)₂ and the like. Another example of an alkyl group as partof a larger structure includes the residue —NHC(O)-alkyl-C(O)OH, whiche.g., may be —NHC(O)CH₂CH₂C(O)OH when alkyl is —CH₂CH₂—.

“Alkenyl” is understood to refer to a group of 2 or more carbon atoms,such as 2 to 10 carbon atoms and more preferably 2 to 6 carbon atoms andhaving at least one and preferably from one to two double bonds.Examples of an alkenyl group include —C═CH₂, —CH₂CH═CHCH₃ and—CH₂CH═CH—CH═CH₂.

“Alkynyl” refers to alkynyl group preferably having from 2 to 10 carbonatoms and more preferably 3 to 6 carbon atoms and having at least onetriple bond, such as the moiety —C≡CH.

“Substituted alkyl” refers to an alkyl group having from 1 to 5substituents. For instance, an alkyl group substituted with a group suchas halo, nitro, cyano, oxo, aryl, alkoxy, alkoxycarbonyl, acyl, acyloxy,acylamino, amino, hydroxyl, carboxyl, carboxylalkyl, thiol, thioalkyl,heterocyclyl, —OS(O)₂-alkyl, and the like is a substituted alkyl. Thesubstituent may be further substituted, for example, by halo, hydroxy,alkyl, alkoxy, aryl, substituted aryl, substituted alkyl, substitutedaralkyl, and the like. Likewise, “substituted alkenyl” and “substitutedalkynyl” refer to alkenyl or alkynyl groups having 1 to 5 substituents.

“Aryl” intends a monocyclic, bicyclic or tricyclic aromatic hydrocarbonring system having 6 to 14 carbon atoms. Examples of groups whoseradicals are aryl groups include, e.g., benzene, naphthalene, indane andtetralin.

“Heteroaryl” refers to an aromatic ring system having at least oneannular heteroatom selected from O, N, or S. An heteroaryl group ispreferably a 5- or 6-membered aromatic ring containing 1-3 annularheteroatoms selected from O, N, or S; a bicyclic 9- or 10-memberedaromatic ring system (meaning the ring system has 9 or 10 annular atoms)containing 1-3 annular heteroatoms selected from O, N, or S; or atricyclic 13- or 14-membered aromatic ring system (meaning the ringsystem has 13 or 14 annular atoms) containing 1-3 annular heteroatomsselected from O, N, or S. Examples of groups whose radicals areheteroaryl groups include e.g., imidazole, pyridine, indole, thiophene,benzopyranone, thiazole, furan, benzimidazole, benzoxazole,benzthiazole, quinoline, isoquinoline, quinoxaline, pyrimidine,pyrazine, tetrazole and pyrazole.

“Substituted aryl” refers to an aryl group having from 1 to 5substituents. For instance, an aryl group substituted with 1 to 5 groupssuch as halo, nitro, cyano, oxo, aryl, alkoxy, alkyl, acyl, acylamino,amino, hydroxyl, carboxyl, carboxylalkyl, thiol, thioalkyl,heterocyclyl, —OS(O)₂-alkyl, and the like is a substituted aryl.Likewise, “substituted heteroaryl” refers to heteroaryl groups having 1to 5 substituents.

“Aralkyl” refers to a residue in which an aryl moiety is attached to theparent structure via an alkyl residue. “Aralkenyl” and “aralkynyl”residues refer to aryl moieties attached to the parent structure via analkenyl or alkynyl residue, respectively. Examples include benzyl(—CH₂-Ph), phenethyl (—CH₂CH₂Ph), phenylvinyl (—CH═CH-Ph), phenylallyland the like.

“Heterocyclyl” or “heterocyclic” refers to a ring system having at leastone annular heteroatom selected from O, N, or S. Examples ofheterocycles whose radicals are heterocyclyl groups includetetrahydropyran, morpholine, pyrrolidine, piperidine, thiazolidine,dioxane, tetrahydrofuran, tetrahydrofuranone and the like. A specificexample of a heterocyclyl residue is tetrahydropyranyl.

“Substituted heterocyclyl” or “substituted heterocyclic” refers to aheterocyclyl group having from 1 to 5 substituents. For instance, aheterocyclyl group substituted with 1 to 5 groups such as halo, nitro,cyano, oxo, aryl, alkoxy, alkyl, acyl, acylamino, amino, hydroxyl,carboxyl, carboxylalkyl, thiol, thioalkyl, heterocyclyl, —OS(O)₂-alkyl,and the like is a substituted heterocyclyl. A particular example of asubstituted heterocyclyl is N-methylpiperazinyl.

“Acyl” refers to and includes the groups —C(O)H, —C(O)alkyl,—C(O)substituted alkyl, —C(O)alkenyl, —C(O)substituted alkenyl,—C(O)alkynyl, —C(O)substituted alkynyl, —C(O)cycloalkyl,—C(O)substituted cycloalkyl, —C(O)aryl, —C(O)substituted aryl,—C(O)heteroaryl, —C(O)substituted heteroaryl, —C(O)heterocyclic, and—C(O)substituted heterocyclic wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclyl and substituted heterocyclyl are as definedherein.

“Halo” refers to fluoro, chloro, bromo or iodo.

“Perhaloalkyl” refers to an alkyl group where each H of the hydrocarbonis replaced with halo. Examples of perhaloalkyl groups include —CF₃,CF₂Cl and —CF₂CF₃ and the like.

“Alkoxy” refers to an alkyl group that is connected to the parentstructure through an oxygen atom (—O-alkyl). When a cycloalkyl group isconnected to the parent structure through an oxygen atom, the group mayalso be referred to as a cycloalkoxy group. Examples include methoxy,ethoxy, propoxy, isopropoxy, cyclopropyloxy, cyclohexyloxy and the like.A “perhaloalkoxy” intends a perhaloalkyl group attached to the parentstructure through an oxygen, such as the residue —O—CF₃.

“Aryloxy” refers to an aryl group that is connected to the parentstructure through an oxygen atom (—O-aryl), which by way of exampleincludes the residues phenoxy, naphthoxy, and the like. “Substitutedaryloxy” refers to a substituted aryl group connected to the parentstructure through an oxygen atom (—O-substituted aryl).

As used herein the term “substituent” or “substituted” means that ahydrogen radical on a compound or group (such as, for example, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, aryl, substituted aryl, aralkyl, substituted aralkyl,heteroaryl, substituted heteroaryl, heteroaralkyl, substitutedheteroaralkyl, cycloalkyl, substituted cycloalkyl, heterocycloalkyl,substituted heterocycloalkyl, heterocyclyl and substituted heterocyclyl)is replaced with any desired group that does not substantially adverselyaffect the stability of the compound. In one embodiment, desiredsubstituents are those which do not adversely affect the activity of acompound. The term “substituted” refers to one or more substituents(which may be the same or different), each replacing a hydrogen atom.Examples of substituents include, but are not limited to, halogen (F,Cl, Br, or I), hydroxyl, amino, alkylamino, arylamino,alkoxycarbonylamino, acylamino, dialkylamino, diarylamino, cyano, nitro,mercapto, oxo (═O), thioxo (═S), or imino (═N-alkyl), formyl, carbamoyl,carboxyl, thioureido, thiocyanato, aminosulfonyl, alkylsulfonyl,arylsulfonyl, alkyl, alkenyl, alkoxy, mercaptoalkoxy, aryl, heteroaryl,cyclyl, heterocyclyl, acyl, acyloxy, alkoxycarbonyl, wherein alkyl,alkenyl, alkoxy, aryl, heteroaryl, cyclyl, and heterocyclyl areoptionally substituted with alkyl, aryl, heteroaryl, halogen, hydroxyl,amino, mercapto, cyano, nitro, oxo, thioxo, or imino. In otherembodiments, substituents on any group (such as, for example, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, aryl, substituted aryl, aralkyl, substituted aralkyl,heteroaryl, substituted heteroaryl, heteroaralkyl, substitutedheteroaralkyl, cycloalkyl, substituted cycloalkyl, heterocycloalkyl,substituted heterocycloalkyl, heterocyclyl and substituted heterocyclyl)can be at any atom of that group (such as on a carbon atom of theprimary carbon chain of a substituted alkyl group or on a substituentalready present on a substituted alkyl group) or at any atom of, whereinany group that can be substituted (such as, for example, alkyl, alkenyl,alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl, cycloalkyl, cyclyl,heterocycloalkyl, and heterocyclyl) can be optionally substituted withone or more substituents (which may be the same or different), eachreplacing a hydrogen atom. Examples of suitable substituents include,but not limited to alkyl, alkenyl, alkynyl, cyclyl, cycloalkyl,heterocyclyl, heterocycloalkyl, aralkyl, heteroaralkyl, aryl,heteroaryl, halogen, haloalkyl, cyano, nitro, alkoxy, aryloxy, hydroxyl,hydroxylalkyl, oxo, carboxyl, formyl, alkylcarbonyl, alkylcarbonylalkyl,alkoxycarbonyl, alkylcarbonyloxy, aryloxycarbonyl, heteroaryloxy,heteroaryloxycarbonyl, thio, mercapto, mercaptoalkyl, arylsulfonyl,amino, aminoalkyl, dialkylamino, alkylcarbonylamino, alkylaminocarbonyl,or alkoxycarbonylamino; alkylamino, arylamino, diarylamino,alkylcarbonyl, or arylamino-substituted aryl; arylalkylamino,aralkylaminocarbonyl, amido, alkylaminosulfonyl, arylaminosulfonyl,dialkylaminosulfonyl, alkylsulfonylamino, arylsulfonylamino, imino,carbamoyl, thioureido, thiocyanato, sulfoamido, sulfonylalkyl,sulfonylaryl, or mercaptoalkoxy. Additional suitable substituents onalkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl,cycloalkyl, cyclyl, heterocycloalkyl, and heterocyclyl.

“Geminal” refers to the relationship between two moieties that areattached to the same atom. For example, in the residue —CH₂—CR′R″—, R′and R″ are geminal and R′ may be referred to as a geminal R group to R″.As an illustrative example, when R′ is taken together with a geminal Rgroup, e.g. R″ in the residue CR′R″ to form a carbonyl moiety, theresidue is C═O.

“Vicinal” refers to the relationship between two moieties that areattached to adjacent atoms. For example, in the residue —CHR′—CHR″—, R′and R″ are vicinal and R′ may be referred to as a vicinal R group to R″.As an illustrative example, when R′ is taken together with a vicinal Rgroup, e.g. R″ in the residue —CHR′—CHR″— to form a bond, the residue is—CH═CH—.

“Pharmaceutically acceptable salt” refers to pharmaceutically acceptablesalts of a compound described herein, such as a compound of Formula (I),(II), (III) or (IV) or other nitroxyl donor of the invention, whichsalts may be derived from organic and inorganic acids, such ashydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate,oxalate and the like; when the molecule contains an acid functionality,salts may be derived from a variety of organic or inorganic counter ionswell known in the art and include, by way of example only, sodium,potassium, calcium, magnesium, ammonium, tetraalkylammonium, and thelike. Illustrative salts include, but are not limited to, sulfate,citrate, acetate, chloride, bromide, iodide, nitrate, bisulfate,phosphate, acid phosphate, lactate, salicylate, acid citrate, tartrate,oleate, tannate, pantothenate, bitartrate, ascorbate, succinate,maleate, besylate, fumarate, gluconate, glucaronate, saccharate,formate, benzoate, glutamate, methane sulfonate, ethanesulfonate,benzenesulfonate, and p-toluenesulfonate salts. Accordingly, a salt maybe prepared from a compound of any one of the formulae disclosed hereinhaving a basic functional group, such as an amino functional group, anda pharmaceutically acceptable inorganic or organic acid. Suitable acidsinclude sulfuric acid, citric acid, acetic acid, hydrochloric acid,hydrogen bromide, hydrogen iodide, nitric acid, phosphoric acid, lacticacid, salicylic acid, tartaric acid, ascorbic acid, succinic acid,maleic acid, besylic acid, fumaric acid, gluconic acid, glucaronic acid,formic acid, benzoic acid, glutamic acid, methanesulfonic acid,ethanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic acid. Asalt may also be prepared from a compound of any one of the formulaedisclosed herein having an acidic functional group, such as a carboxylicacid functional group, and a pharmaceutically acceptable inorganic ororganic base. Suitable bases include, but are not limited to, hydroxidesof alkali metals such as sodium, potassium, and lithium; hydroxides ofalkaline earth metal such as calcium and magnesium; hydroxides of othermetals, such as aluminum and zinc; ammonia, and organic amines, such asunsubstituted or hydroxy-substituted mono-, di-, or trialkylamines;dicyclohexylamine; tributylamine; pyridine; N-methyl-N-ethylamine;diethylamine; triethylamine; mono-, bis-, or tris-(2-hydroxy-lower alkylamines), such as mono-, bis-, or tris(2-hydroxyethyl)amine, ortris-(hydroxymethyl)methylamine, N,N-di-lower alkyl-N-(hydroxy loweralkyl)-amines, such as N,N-dimethyl-N-(2-hydroxyethyl)amine, orN-methyl-D-glucamine; and amino acids such as arginine, lysine, and thelike.

Unless clearly indicated otherwise, “an individual” as used hereinintends a mammal, including but not limited to a human. For example, anindividual may have or is suspected of having a disease or conditionthat are responsive to nitroxyl therapy, including heart failure,ischemia/reperfusion injury and cancer.

The term “effective amount” intends such amount of a compound or apharmaceutically acceptable salt thereof, which in combination with itsparameters of efficacy and toxicity, as well as based on the knowledgeof the practicing specialist should be effective in a given therapeuticform. As is understood in the art, an effective amount may be in one ormore doses.

As used herein, “treatment” or “treating” is an approach for obtaining abeneficial or desired result, including clinical results. For purposesof this invention, beneficial or desired results include but are notlimited to inhibiting and/or suppressing the onset and/or development ofa disease or condition that is responsive to nitroxyl therapy orreducing the severity of such disease or condition, such as reducing thenumber and/or severity of symptoms associated with the disease orcondition, increasing the quality of life of those suffering from thedisease or condition, decreasing the dose of other medications requiredto treat the disease or condition, enhancing the effect of anothermedication an individual is taking for the disease or condition andprolonging survival of individuals having the disease or condition. Thedisease or condition may be a cardiovascular disease or condition, whichincludes, but is not limited to, coronary obstructions, coronary arterydisease (CAD), angina, heart attack, myocardial infarction, high bloodpressure, ischemic cardiomyopathy and infarction, diastolic heartfailure, pulmonary congestion, pulmonary edema, cardiac fibrosis,valvular heart disease, pericardial disease, circulatory congestivestates, peripheral edema, ascites, Chagas' disease, ventricularhypertrophy, heart valve disease, heart failure, including but notlimited to congestive heart failure such as acute congestive heartfailure and acute decompensated heart failure. Related symptoms that maybe alleviated by the methods herein include shortness of breath,fatigue, swollen ankles or legs, angina, loss of appetite, weight gainor loss, associated with aforementioned diseases or disorders. Thedisease or condition may involve ischemia/reperfusion injury. Thedisease or condition may be a cancerous disease or condition, whichincludes, but is not limited to, breast, prostate, pancreatic orcolorectal cancer.

As used herein, “preventing” refers to reducing the probability ofdeveloping a disorder or condition in an individual who does not have,but is at risk of developing a disorder or condition.

An individual “at risk” may or may not have a detectable disease orcondition, and may or may not have displayed a detectable disease orcondition prior to the treatment methods described herein. “At risk”denotes that an individual has one or more so-called risk factors, whichare measurable parameters that correlate with development of a diseaseor condition and are known in the art. An individual having one or moreof these risk factors has a higher probability of developing the diseaseor condition than an individual without these risk factor(s). Forexample, an individual with a family history of heart disease,uncontrolled hypertension (high blood pressure), physical inactivity,obesity (more than 20% over one's ideal body weight) or uncontrolleddiabetes may be at risk for heart diseases.

“Nitroxyl” refers to the species HNO.

As used herein, a compound is a “nitroxyl donor” if it donates nitroxylunder physiological conditions. Preferably, the nitroxyl donor iscapable of donating an effective amount of nitroxyl in vivo and has asafety profile indicating the compound would be tolerated by anindividual in the amount necessary to achieve a therapeutic effect. Oneof ordinary skill in the art would be able to determine the safety ofadministering particular compounds and dosages to live subjects. Oneskilled in the art may also determine whether a compound is a nitroxyldonor by evaluating whether it releases HNO under physiologicalconditions. Compounds are easily tested for nitroxyl donation withroutine experiments. Although it is impractical to directly measurewhether nitroxyl is donated, several tests are accepted for determiningwhether a compound donates nitroxyl. For example, the compound ofinterest can be placed in solution, for example in water, in a sealedcontainer. After sufficient time for disassociation has elapsed, such asfrom several minutes to several hours, the headspace gas is withdrawnand analyzed to determine its composition, such as by gas chromatographyand/or mass spectroscopy. If the gas N₂O is formed (which occurs by HNOdimerization, see Smith, P. A. S, and Hein, G. E. J. Am. Chem. Soc.1960, 82, 5731-5740; and Kohout, F. C. and Lampe, F. W. J. Am. Chem.Soc. 1965, 87, 5795-5796), the test is positive for nitroxyl donationand the compound is a nitroxyl donor. The level of nitroxyl donatingability may be expressed as a percentage of a compound's theoreticalmaximum. A compound that donates a “significant level of nitroxyl”intends a compound that donates 40% or more or 50% or more of itstheoretical maximum amount of nitroxyl. In one variation, the compoundsfor use herein donate 60% or more of the theoretical maximum amount ofnitroxyl. In another variation, the compounds for use herein donate 70%or more of the theoretical maximum amount of nitroxyl. In anothervariation, the compounds for use herein donate 80% or more of thetheoretical maximum amount of nitroxyl. In another variation, thecompounds for use herein donate 90% or more of the theoretical maximumamount of nitroxyl. In yet another variation, the compounds for useherein donate between about 70% and about 90% of the theoretical maximumamount of nitroxyl. In yet another variation, the compounds for useherein donate between about 85% and about 95% of the theoretical maximumamount of nitroxyl. In yet another variation, the compounds for useherein donate between about 90% and about 95% of the theoretical maximumamount of nitroxyl. Compounds that donate less than 40% or less than 50%of their theoretical amount of nitroxyl are still nitroxyl donors andmay be used in the invention disclosed herein. A compound that donatesless than 50% of the theoretical amount of nitroxyl may be used in themethods described, and may require higher dosing levels as compared tocompounds that donate a significant level of nitroxyl. Alternatively,HNO formation from compounds of the invention can be assessed by theability of the compounds to reductively nitrosylate ferric heme groupsyielding the relatively stable ferrous nitrosyl complexes as judged byultraviolet/visible (UV/Vis) and Electron Paramagnetic Resonance (EPR)spectroscopies (Sha, X. et al J. Am. Chem. Soc. 2006, 128, 9687-9692).Nitroxyl donation also can be detected by exposing the test compound tometmyoglobin (Mb³⁺). Nitroxyl reacts with Mb³⁺ to form an Mb²⁺-NOcomplex, which can be detected by changes in the ultraviolet/visiblespectrum or by Electron Paramagnetic Resonance (EPR). The Mb²⁺-NOcomplex has an EPR signal centered around a g-value of about 2. Nitricoxide, on the other hand, reacts with Mb³⁺ to form an Mb³⁺-NO complexthat is EPR silent. Accordingly, if the candidate compound reacts withMb³⁺ to form a complex detectable by common methods such asultraviolet/visible or EPR, then the test is positive for nitroxyldonation. Testing for nitroxyl donation may be performed atphysiologically relevant pH.

A “positive inotrope” as used herein is an agent that causes an increasein myocardial contractile function. Such an agent includes abeta-adrenergic receptor agonist, an inhibitor of phosphodiesteraseactivity, and calcium-sensitizers. Beta-adrenergic receptor agonistsinclude, among others, dopamine, dobutamine, terbutaline, andisoproterenol. Analogs and derivatives of such compounds are alsointended. For example, U.S. Pat. No. 4,663,351 describes a dobutamineprodrug that can be administered orally. One of ordinary skill in theart would be able to determine if a compound is capable of causingpositive inotropic effects and also additional beta-agonist compounds.In particular embodiments, the beta-receptor agonist is selective forthe beta-1 receptor. However, in other embodiments the beta-agonist isselective for the beta-2 receptor, or is not selective for anyparticular receptor.

Diseases or conditions that are “responsive to nitroxyl therapy” intendsany disease or condition in which administration of a compound thatdonates an effective amount of nitroxyl under physiological conditionstreats and/or prevents the disease or condition, as those terms aredefined herein. A disease or condition whose symptoms are suppressed ordiminished upon administration of nitroxyl donor is a disease orcondition responsive to nitroxyl therapy. Non-limiting examples ofdiseases or conditions that are responsive to nitroxyl therapy includecoronary obstructions, coronary artery disease (CAD), angina, heartattack, myocardial infarction, high blood pressure, ischemiccardiomyopathy and infarction, diastolic heart failure, pulmonarycongestion, pulmonary edema, cardiac fibrosis, valvular heart disease,pericardial disease, circulatory congestive states, peripheral edema,ascites, Chagas' disease, ventricular hypertrophy, heart valve disease,heart failure, including but not limited to congestive heart failuresuch as acute congestive heart failure and acute decompensated heartfailure. Other cardiovascular diseases or conditions are also intended,as are diseases or conditions that implicate ischemia/reperfusioninjury. Cancer is another example of disease or condition that isresponsive to nitroxyl therapy.

Nitroxyl Donor Compounds

The compounds disclosed herein are a novel class of nitroxyl donors thatrelease HNO under physiological conditions.

Preferably, a compound of this invention releases efficacious amounts ofHNO at a controlled rate under physiological conditions. For example,the rate of HNO release from an acyloxy nitroso compound of thisinvention may be modulated by varying the nature of the acyloxy groupand the structure bearing the nitroso moiety.

In one embodiment, the invention embraces a compound of the formula (I):

where each R¹ and R² is independently a substituted or unsubstitutedC₁-C₈ alkyl, substituted or unsubstituted C₂-C₈ alkenyl, substituted orunsubstituted C₂-C₈ alkynyl, or R¹ and R² are taken together to form anunsubstituted or substituted 6 or 7-membered carbocyclic moiety or anunsubstituted or substituted 5, 6 or 7-membered heterocyclic moiety;

D is selected from the group consisting of alkyl-C(O)—, substitutedalkyl-C(O)—, perhaloalkyl-C(O)—, alkenyl-C(O)—, substitutedalkenyl-C(O)—, alkynyl-C(O)—, substituted alkynyl-C(O)—, aryl-C(O)—,substituted aryl-C(O)—, heteroaryl-C(O)—, substituted heteroaryl-C(O)—,heterocyclyl-C(O)— and —P(O)(OC₁-C₈alkyl)₂;

provided that the compound is other than 1-nitrosocycloheptyl acetate,1-nitrosocycloheptyl benzoate, 9-nitrosobicyclo[3.3.1]nonan-9-yl acetateor 8-methyl-3-nitroso-8-azabicyclo[3.2.1]octan-3-yl acetate; and (i)when R¹ or R² is an unsubstituted C₁-C₈ alkyl, the R¹ or R² that is anunsubstituted C₁-C₈ alkyl is other than methyl or propyl and D is otherthan an NSAID moiety; (ii) when R¹ and R² are taken together to form anunsubstituted 6-membered carbocyclic moiety, D is other thann-alkyl-C(O)—, ClCH₂—C(O)—, CCl₃—C(O)—, CF₃—C(O)—, (CH₃)₃C—C(O)—,alkenyl-C(O)—, substituted alkenyl-C(O)—, unsubstituted aryl-C(O)—,mono-substituted aryl-C(O)— or an NSAID moiety; (iii) when R¹ and R² aretaken together to form a substituted 6-membered carbocyclic moiety, thesubstituted 6-membered carbocyclic moiety is a monocyclic or bicyclicring that is substituted with a moiety other than alkyl, nitroso, acyl,oxime, and substituted alkenyl; or (iv) when R¹ and R² are takentogether to form an unsubstituted or substituted 5 or 6-memberedheterocyclic moiety, the 5 or 6-membered heterocyclic moiety is amonocyclic or bicyclic ring other than dioxane or acyloxy-substitutedtetrahydropyan;

or a salt or solvate thereof.

In another embodiment, the compound is of the formula (I) where each R¹and R² is independently a substituted C₁-C₈ alkyl. In one embodiment,the compound is of the formula (I) where each R¹ and R² is independentlya moiety of the formula C₁-C₄alkyl-O—C₁-C₄alkyl- or acyl-O—C₁-C₄alkyl-.In one specific embodiment, the compound of the formula (I), where R¹and R² are each of the formula CH₃CH₂—O—CH₂— or CH₃C(O)—O—CH₂—. In amore specific embodiment, the compound of the formula (I), where R¹ andR² are each of the formula CH₃CH₂—O—CH₂— or CH₃C(O)—O—CH₂— and D isalkyl-C(O)—, e.g. acetyl.

In one embodiment, the invention embraces a compound of the formula (I),where D is selected from the group consisting of alkyl-C(O)—,substituted alkyl-C(O)—, perhaloalkyl-C(O)—, alkenyl-C(O)—, substitutedalkenyl-C(O)—, alkynyl-C(O)—, substituted alkynyl-C(O)—, aryl-C(O)—,substituted aryl-C(O)—, heteroaryl-C(O)—, substituted heteroaryl-C(O)—,heterocyclic-C(O)—. In one variation, the compound is of formula (I),where D is as described in this paragraph and each R¹ and R² isindependently a substituted C₁-C₈ alkyl. In another variation of thisembodiment, each R¹ and R² is independently a moiety of the formulaC₁-C₄alkyl-O—C₁-C₄alkyl- or acyl-O—C₁-C₄alkyl-. In one specificvariation, R¹ and R² are each of the formula CH₃CH₂—O—CH₂— orCH₃C(O)—O—CH₂—.

In one embodiment, the invention embraces a compound of the formula (I),where D is unsubstituted C₁-C₄alkyl-C(O)—, perhaloalkyl-C(O)—,substituted aryl-C(O)—, substituted aralkyl-C(O)—, or substitutedC₁-C₄alkyl-C(O)— where the substitution is one or more substituentsselected from the group consisting of halo, cyano, alkoxy, acyloxy,substituted acyloxy [e.g. CH₃OCH₂CH₂OCH₂C(O)O—], acylamino, substitutedacylamino, alkylamino, substituted alkylamino, dialkylamino,N-acyl-substituted alkylamino [e.g. (AcOCH₂)₂CHN(Ac)—],N-alkyl-substituted alkylamino [e.g. (AcOCH₂)₂CHN(Me)-],alkoxycarbonylamino [e.g. t-BuOC(O)NH—], substituted alkoxycarbonylamino[e.g. PhCH₂OC(O)NH—], alkoxycarbonyl, heterocyclyl and substitutedheterocyclyl. In one variation, D is a di-substituted C₁-C₄alkyl-C(O)—where the C₁-C₄alkyl-C(O)— is of the formula

In one variation, the compound is of formula (I), where D is asdescribed in this paragraph and each R¹ and R² is independently asubstituted C₁-C₈ alkyl. In another variation of this embodiment, eachR¹ and R² is independently a moiety of the formulaC₁-C₄alkyl-O—C₁-C₄alkyl- or acyl-O—C₁-C₄alkyl-. In one specificvariation, R¹ and R² are each of the formula CH₃CH₂—O—CH₂— orCH₃C(O)—O—CH₂—.

In one embodiment, the invention embraces a compound of the formula (I),where D is C₁-C₈alkyl-C(O)—. In one variation of this embodiment, eachR¹ and R² is independently a substituted C₁-C₈ alkyl. In anothervariation of this embodiment, each R¹ and R² is independently a moietyof the formula C₁-C₄alkyl-O—C₁-C₄alkyl- or acyl-O—C₁-C₄alkyl-. In onespecific variation, the compound is of formula (I), where D isC₁-C₈alkyl-C(O)— and R¹ and R² are each of the formula CH₃CH₂—O—CH₂— orCH₃C(O)—O—CH₂—.

In one embodiment, the invention embraces a compound of the formula (I),where D is a moiety of the formula:

In another embodiment, the compound is of the formula (I), where D is amoiety of the formula:

In one embodiment, the invention embraces a compound of the formula (I),where D is CH₃—C(O)—. In another embodiment, the compound is of theformula (I), where D is P(O)(OC₁-C₈alkyl)₂.

In another embodiment, the invention embraces a compound of the formula(Ia):

where R³ is unsubstituted or substituted alkyl, perhaloalkyl,unsubstituted or substituted alkenyl, unsubstituted or substitutedalkynyl, unsubstituted or substituted aryl, unsubstituted or substitutedheteroaryl or heterocyclyl. R¹ and R² of formula (Ia) are as defined forformula (I).

In yet another embodiment, the compound is of the formula (Ia), where R³is C₁-C₈alkyl. In one specific embodiment, the compound is of theformula (Ia), where R³ is methyl.

In one embodiment, the invention embraces a compound of the formula(II):

where D is selected from the group consisting of alkyl-C(O)—,substituted alkyl-C(O)—, perhaloalkyl-C(O)—, alkenyl-C(O)—, substitutedalkenyl-C(O)—, alkynyl-C(O)—, substituted alkynyl-C(O)—, aryl-C(O)—,substituted aryl-C(O)—, heteroaryl-C(O)—, substituted heteroaryl-C(O)—,heterocyclyl-C(O)— and —P(O)(OC₁-C₈alkyl)₂;

X is O, NR⁴, CR⁵R⁶, S, S(O) or S(O)₂;

Y is CR⁵R⁶ or CR⁵R⁶—CR⁷R⁸;

Z is CR⁵R⁶ or a bond, provided that when X is CR⁵R⁶, Z is CR⁵R⁶;

R⁴ is H, substituted or unsubstituted C₁-C₈ alkyl, substituted orunsubstituted C₂-C₈ alkenyl, substituted or unsubstituted C₂-C₈ alkynyl,substituted or unsubstituted acyl, alkoxycarbonyl, substituted orunsubstituted heterocyclyl, substituted or unsubstituted aryl,substituted or unsubstituted aralkyl, or sulfonyl;

each R⁵, R⁶, R⁷, R⁸, R^(11a), R^(11b), R^(12a), and R^(12b) isindependently H, substituted or unsubstituted C₁-C₈ alkyl, halo,hydroxyl, alkoxy, cyano, nitro, or is taken together with a geminal Rgroup to form a carbonyl moiety, or is taken together with a vicinal Rgroup to form a bond, or is taken together with another R group to forma ring;

provided that the compound is other than 1-nitrosocycloheptyl acetate,1-nitrosocycloheptyl benzoate, 9-nitrosobicyclo[3.3.1]nonan-9-yl acetateor 8-methyl-3-nitroso-8-azabicyclo[3.2.1]octan-3-yl acetate; and (i)when X, Y, Z, R^(11a), R^(11b), R^(12a), and R^(12b) are taken togetherto form an unsubstituted 6-membered carbocyclic moiety, D is other thann-alkyl-C(O)—, ClCH₂—C(O)—, CCl₃-C(O)—, CF₃—C(O)—, (CH₃)₃C—C(O)—,alkenyl-C(O)—, substituted alkenyl-C(O)—, unsubstituted aryl-C(O)—,mono-substituted aryl-C(O)— or an NSAID moiety; (ii) when X, Y, Z,R^(11a), R^(11b), R^(12a), and R^(12b) are taken together to form asubstituted 6-membered carbocyclic moiety, the substituted 6-memberedcarbocyclic moiety is a monocyclic or bicyclic ring that is substitutedwith a moiety other than alkyl, nitroso, acyl, oxime, and substitutedalkenyl; or (iii) when X, Y, Z, R^(11a), R^(11b), R^(12a), and R^(12b)are taken together to form an unsubstituted or substituted 5 or6-membered heterocyclic moiety, the 5 or 6-membered heterocyclic moietyis a monocyclic or bicyclic ring other than dioxane oracyloxy-substituted tetrahydropyran;

or a salt or solvate thereof.

In another embodiment, the compound is of the formula (II), where D isselected from the group consisting of alkyl-C(O)—, substitutedalkyl-C(O)—, perhaloalkyl-C(O)—, alkenyl-C(O)—, substitutedalkenyl-C(O)—, alkynyl-C(O)—, substituted alkynyl-C(O)—, aryl-C(O)—,substituted aryl-C(O)—, heteroaryl-C(O)—, substituted heteroaryl-C(O)—,heterocyclic-C(O)—.

In one embodiment, the invention embraces a compound of the formula(II), where D is unsubstituted C₁-C₄alkyl-C(O)—, perhaloalkyl-C(O)—,substituted aryl-C(O)—, substituted aralkyl-C(O)—, or substitutedC₁-C₄alkyl-C(O)— where the substitution is one or more substituentsselected from the group consisting of halo, cyano, alkoxy, acyloxy,substituted acyloxy [e.g. CH₃OCH₂CH₂OCH₂C(O)O—], acylamino, substitutedacylamino, alkylamino, substituted alkylamino, dialkylamino,N-acyl-substituted alkylamino [e.g. (AcOCH₂)₂CHN(Ac)—],N-alkyl-substituted alkylamino [e.g. (AcOCH₂)₂CHN(Me)-],alkoxycarbonylamino [e.g. t-BuOC(O)NH—], substituted alkoxycarbonylamino[e.g. PhCH₂OC(O)NH—], alkoxycarbonyl, heterocyclyl and substitutedheterocyclyl. In one variation, D is a di-substituted C₁-C₄alkyl-C(O)—where the C₁-C₄alkyl-C(O)— moiety is of the formula

In another embodiment, the invention embraces a compound of the formula(II), where D is a moiety of the formula:

In another embodiment, the compound is of the formula (II), where D is amoiety of the formula:

In another embodiment, the compound is of the formula (II), where D isP(O)(OC₁-C₈alkyl)₂. In one embodiment, the compound is of the formula(II), where D is selected from P(O)(OCH₂CH₃)₂ and P(O)(OCH₂CH₂CH₂CH₃)₂.

In some embodiments, the compound is of the formula (II) or anyvariations described herein, where X, Y, Z, R^(11a), R^(11b), R^(12a),and R^(12b) are taken together to form a moiety of the structureselected from:

In one variation, the compound is of the formula (II) where X is O and Yand Z are each CR⁵R⁶ where each R⁵ and R⁶ is independently H,substituted or unsubstituted C₁-C₈ alkyl, halo, hydroxyl, alkoxy, cyano,nitro, or is taken together with a geminal R group to form a carbonylmoiety. In one such variation, each Y and Z is CH₂. In a specificvariation, each Y and Z is CH₂ and each R^(11a), R^(11b), R^(12a) andR^(12b) is H. In one specific variation, the compound is of the formula(II), where X is O, each Y and Z is CH₂, each R^(11a), R^(11b), R^(12a)and R^(12b) is H, and D is selected from the group consisting ofalkyl-C(O)—, substituted alkyl-C(O)—, perhaloalkyl-C(O)—, alkenyl-C(O)—,substituted alkenyl-C(O)—, alkynyl-C(O)—, substituted alkynyl-C(O)—,aryl-C(O)—, substituted aryl-C(O)—, heteroaryl-C(O)—, substitutedheteroaryl-C(O)— and heterocyclyl-C(O)—.

In another variation, the compound is of the formula (II) where X isNR⁴, and Y and Z are each CR⁵R⁶ where each R⁵ and R⁶ is independently H,substituted or unsubstituted C₁-C₈ alkyl, halo, hydroxyl, alkoxy, cyano,nitro, or is taken together with a geminal R group to form a carbonylmoiety. In one such variation, X is NR⁴ where R⁴ is substituted orunsubstituted C₁-C₈ alkyl (e.g. methyl) or substituted or unsubstitutedacyl (e.g. acetyl or benzoyl). In another such variation, each R⁵ and R⁶is independently H, or C₁-C₈ alkyl (e.g. methyl). In yet another suchvariation, R^(11a), R^(11b), R^(12a) and R^(12b) is H. In one specificvariation, the compound is of the formula (II) where X is NR⁴ where R⁴is substituted or unsubstituted C₁-C₈ alkyl or substituted orunsubstituted acyl, Y and Z are each CR⁵R⁶ where each R⁵ and R⁶ isindependently H, or C₁-C₈ alkyl, each R^(11a), R^(11b), R^(12a) andR^(12b) is H and D is selected from the group consisting of alkyl-C(O)—,substituted alkyl-C(O)—, perhaloalkyl-C(O)—, alkenyl-C(O)—, substitutedalkenyl-C(O)—, alkynyl-C(O)—, substituted alkynyl-C(O)—, aryl-C(O)—,substituted aryl-C(O)—, heteroaryl-C(O)—, substituted heteroaryl-C(O)—,heterocyclyl-C(O)— and —P(O)(OC₁-C₈alkyl)₂

In another variation, the compound is of the formula (II) where X, Y andZ are each CR⁵R⁶ where each R⁵ and R⁶ is independently H, substituted orunsubstituted C₁-C₈ alkyl, halo, hydroxyl, alkoxy, cyano, nitro, or istaken together with a geminal R group to form a carbonyl moiety. In onesuch variation, each R⁵ and R⁶ is H. In another such variation, eachR^(11a), R^(11b), R^(12a) and R^(12b) is H. In one specific variation,the compound is of the formula (II) where X, Y and Z are each CH₂, eachR^(11a), R^(11b), R^(12a) and R^(12b) is H and D is selected from thegroup consisting of alkyl-C(O)—, substituted alkyl-C(O)—,perhaloalkyl-C(O)—, alkenyl-C(O)—, substituted alkenyl-C(O)—,alkynyl-C(O)—, substituted alkynyl-C(O)—, aryl-C(O)—, substitutedaryl-C(O)—, heteroaryl-C(O)—, substituted heteroaryl-C(O)—,heterocyclyl-C(O)— and —P(O)(OC₁-C₈alkyl)₂; provided that D is otherthan n-alkyl-C(O)—, ClCH₂—C(O)—, CCl₃—C(O)—, CF₃—C(O)—, (CH₃)₃C—C(O)—,alkenyl-C(O)—, substituted alkenyl-C(O)—, unsubstituted aryl-C(O)—,mono-substituted aryl-C(O)— or an NSAID moiety.

In yet another variation, the compound is of the formula (II) where X isS, S(O) or S(O)₂, Z is a bond and Y is CR⁵R⁶ where each R⁵ and R⁶ isindependently H, substituted or unsubstituted C₁-C₈ alkyl, halo,hydroxyl, alkoxy, cyano, nitro, or is taken together with a geminal Rgroup to form a carbonyl moiety. In one such variation, Y is CH₂. Inanother such variation, each R^(11a), R^(11b), R^(12a) and R^(12b) is H.In one specific variation, the compound is of the formula (II) where Xis S, Y is CH₂, Z is a bond, each R^(11a), R^(11b), R^(12a) and R^(12b)is H and D is selected from the group consisting of alkyl-C(O)—,substituted alkyl-C(O)—, perhaloalkyl-C(O)—, alkenyl-C(O)—, substitutedalkenyl-C(O)—, alkynyl-C(O)—, substituted alkynyl-C(O)—, aryl-C(O)—,substituted aryl-C(O)—, heteroaryl-C(O)—, substituted heteroaryl-C(O)—,heterocyclyl-C(O)— and —P(O)(OC₁-C₈alkyl)₂.

In one embodiment, the invention embraces a compound of the formula(IIa):

where R³ is unsubstituted or substituted alkyl, perhaloalkyl,unsubstituted or substituted alkenyl, unsubstituted or substitutedalkynyl, unsubstituted or substituted aryl, unsubstituted or substitutedheteroaryl or heterocyclyl;

X is O, NR⁴, CR⁵R⁶, S, S(O) or S(O)₂;

Y is CR⁵R⁶ or CR⁵R⁶—CR⁷R⁸;

Z is CR⁵R⁶ or a bond, provided that when X is CR⁵R⁶, Z is CR⁵R⁶;

R⁴ is H, substituted or unsubstituted C₁-C₈ alkyl, substituted orunsubstituted C₂-C₈ alkenyl, substituted or unsubstituted C₂-C₈ alkynyl,substituted or unsubstituted acyl, alkoxycarbonyl, substituted orunsubstituted heterocyclyl, substituted or unsubstituted aryl,substituted or unsubstituted aralkyl, or sulfonyl;

each R⁵, R⁶, R⁷, R⁸, R^(11a), R^(11b), R^(12a), and R^(12b) isindependently H, substituted or unsubstituted C₁-C₈ alkyl, halo,hydroxyl, alkoxy, cyano, nitro, or is taken together with a geminal Rgroup to form a carbonyl moiety, or is taken together with a vicinal Rgroup to form a bond, or is taken together with another R group to forma ring;

provided that the compound is other than 1-nitrosocycloheptyl acetate,1-nitrosocycloheptyl benzoate, 9-nitrosobicyclo[3.3.1]nonan-9-yl acetateor 8-methyl-3-nitroso-8-azabicyclo[3.2.1]octan-3-yl acetate; and (i)when X, Y, Z, R^(11a), R^(11b), R^(12a), and R^(12b) are taken togetherto form an unsubstituted 6-membered carbocyclic moiety, D is other thann-alkyl-C(O)—, ClCH₂—C(O)—, CCl₃-C(O)—, CF₃—C(O)—, (CH₃)₃C—C(O)—,alkenyl-C(O)—, substituted alkenyl-C(O)—, unsubstituted aryl-C(O)—,mono-substituted aryl-C(O)— or an NSAID moiety; (ii) when X, Y, Z,R^(11a), R^(11b), R^(12a), and R^(12b) b are taken together to form asubstituted 6-membered carbocyclic moiety, the substituted 6-memberedcarbocyclic moiety is a monocyclic or bicyclic ring that is substitutedwith a moiety other than alkyl, nitroso, acyl, oxime, and substitutedalkenyl; or (iii) when X, Y, Z, R^(11a), R^(11b), R^(12a), and R^(12b)are taken together to form an unsubstituted or substituted 5 or6-membered heterocyclic moiety, the 5 or 6-membered heterocyclic moietyis a monocyclic or bicyclic ring other than dioxane oracyloxy-substituted tetrahydropyran;

or a salt or solvate thereof.

In one embodiment, the compound is of the formula (II) or formula (Ia),where X is O, NR⁴, S, S(O) or S(O)₂. In one variation, X is O. Inanother variation, X is S. In yet another variation, X is NR⁴, such aswhere X is an N-unsubstituted C₁-C₈alkyl, N-carbonyl-C₁-C₄alkyl orN-carbonyl-aryl.

In one embodiment, the invention embraces a compound of the formula(III):

where D is selected from the group consisting of alkyl-C(O)—,substituted alkyl-C(O)—, perhaloalkyl-C(O)—, alkenyl-C(O)—, substitutedalkenyl-C(O)—, alkynyl-C(O)—, substituted alkynyl-C(O)—, aryl-C(O)—,substituted aryl-C(O)—, heteroaryl-C(O)—, substituted heteroaryl-C(O)—,heterocyclyl-C(O)— and —P(O)(OC₁-C₈alkyl)₂;

X is O, NR⁴, CR⁵R⁶, S, S(O) or S(O)₂;

R⁴ is H, substituted or unsubstituted C₁-C₈ alkyl, substituted orunsubstituted C₂-C₈ alkenyl, substituted or unsubstituted C₂-C₈ alkynyl,substituted or unsubstituted acyl, alkoxycarbonyl, substituted orunsubstituted heterocyclyl, substituted or unsubstituted aryl,substituted or unsubstituted aralkyl, or sulfonyl; and

each R⁵, R⁶, R^(11a), R^(11b), R^(12a), R^(12b), R^(13a), R^(13b),R^(14a) and R^(14b) is independently H, substituted or unsubstitutedC₁-C₈ alkyl, halo, hydroxyl, alkoxy, cyano or nitro, or is takentogether with a geminal R group to form a carbonyl moiety, or is takentogether with a vicinal R group to form a bond when X is CR⁵R⁶, or istaken together with another R group to form a ring;

provided that the compound is other than9-nitrosobicyclo[3.3.1]nonan-9-yl acetate or8-methyl-3-nitroso-8-azabicyclo[3.2.1]octan-3-yl acetate; and (i) whenX, R^(11a), R^(11b), R^(12a), R^(12b), R^(13a), R^(13b), R^(14a) andR^(14b) are taken together to form an unsubstituted 6-memberedcarbocyclic moiety, D is other than n-alkyl-C(O)—, ClCH₂—C(O)—,CCl₃-C(O)—, CF₃—C(O)—, (CH₃)₃C—C(O)—, alkenyl-C(O)—, substitutedalkenyl-C(O)—, unsubstituted aryl-C(O)—, mono-substituted aryl-C(O)— oran NSAID moiety; (ii) when X, R^(11a), R^(11b), R^(12a), R^(12b),R^(13a), R^(13b), R^(14a) and R^(14b) are taken together to form asubstituted 6-membered carbocyclic moiety, the substituted 6-memberedcarbocyclic moiety is a monocyclic or bicyclic ring that is substitutedwith a moiety other than alkyl, nitroso, acyl, oxime, and substitutedalkenyl; or (iii) when X, R^(11a), R^(11b), R^(12a), R^(12b), R^(13a),R^(13b), R^(14a) and R^(14b) are taken together to form an unsubstitutedor substituted 5 or 6-membered heterocyclic moiety, the 5 or 6-memberedheterocyclic moiety is a monocyclic or bicyclic ring other than dioxaneor acyloxy-substituted tetrahydropyran; or a salt or solvate thereof.

In another embodiment, the compound is of the formula (III), where X isO, NR⁴, S, S(O) or S(O)₂. In one variation, X is O. In anothervariation, X is NR⁴, such as where X is an N-unsubstituted C₁-C₈alkyl,N-carbonyl-C₁-C₄alkyl or N-carbonyl-aryl.

In one embodiment, the invention embraces a compound of the formula(IV):

where X is O, NR⁴, S, S(O) or S(O)₂;

R⁴ is H, substituted or unsubstituted C₁-C₈ alkyl, substituted orunsubstituted C₂-C₈ alkenyl, substituted or unsubstituted C₂-C₈ alkynyl,substituted or unsubstituted acyl, alkoxycarbonyl, substituted orunsubstituted heterocyclyl, substituted or unsubstituted aryl,substituted or unsubstituted aralkyl, or sulfonyl; and

each R⁵, R⁶, R^(11a), R^(11b), R^(12a), R^(12b), R^(13a) and R^(13b) isindependently H, substituted or unsubstituted C₁-C₈ alkyl, halo,hydroxyl, alkoxy, cyano or nitro, or is taken together with a geminal Rgroup to form a carbonyl moiety, or is taken together with a vicinal Rgroup to form a bond when X is CR⁵R⁶, or is taken together with anotherR group to form a ring;

or a salt or solvate thereof. In formula (IV), D is as defined forformula (I) or any variation thereof.

In some embodiments, the compound is of the formula (IV) where at leastone, two, three, four, five or six of R^(11a), R^(11b), R^(12a),R^(12b), R^(13a) and R^(13b) are H. In some embodiments, the compound isof the formula (IV) where one, two, three, four, five or six of R^(11a),R^(11b), R^(12a), R^(12b), R^(13a) and R^(13b) are H. In a specificembodiment, the compound is of the formula (IV) where each R^(11a),R^(11b), R^(12a), R^(12b), R^(13a) and R^(13b) is H.

In another embodiment, the compound is of the formula (IV) where X is S.In some variations of this embodiment, at least one, two, three, four,five or six of R^(11a), R^(11b), R^(12a), R^(12b), R^(13a) and R^(13b)is H. In some variations of this embodiment, one, two, three, four, fiveor six of R^(11a), R^(11b), R^(12a), R^(12b), R^(13a) and R^(13b) is H.In one particular variation of this embodiment, each R^(11a), R^(11b),R^(12a), R^(12b), R^(13a) and R^(13b) is H.

In one embodiment, the invention embraces a compound of the formula (V):

where D is selected from the group consisting of alkyl-C(O)—,substituted alkyl-C(O)—, perhaloalkyl-C(O)—, alkynyl-C(O)—, substitutedalkynyl-C(O)—, substituted aryl-C(O)—, heteroaryl-C(O)—, substitutedheteroaryl-C(O)—, heterocyclyl-C(O)— and —P(O)(OC₁-C₈alkyl)₂, providedthat D is other than n-alkyl-C(O)—, ClCH₂—C(O)—, CCl₃—C(O)—, CF₃—C(O)—,(CH₃)₃C—C(O)—, mono-substituted aryl-C(O)— or an NSAID moiety; and

each R⁵, R⁶, R^(11a), R^(11b), R^(12a), R^(12b), R^(13a), R^(13b),R^(14a) and R^(14b) is independently H, substituted or unsubstitutedC₁-C₈ alkyl, halo, hydroxyl, alkoxy, cyano or nitro, or is takentogether with a geminal R group to form a carbonyl moiety, or is takentogether with a vicinal R group to form a bond, or is taken togetherwith another R group to form a ring;

or a salt or solvate thereof.

In some embodiments, the compound is of the formula (V), where at leastone, two, three, four, five, six, seven, eight, nine or ten of R⁵, R⁶,R^(11a), R^(11b), R^(12a), R^(12b), R^(13a), R^(13b), R^(14a) andR^(14b) are H. In some embodiments, the compound is of the formula (V)where one, two, three, four, five, six, seven, eight, nine or ten of R⁵,R⁶, R^(11a), R^(11b), R^(12a), R^(12b), R^(13a), R^(13b), R^(14a) andR^(14b) are H. In one specific embodiment, the compound is of theformula (V), where each R⁵, R⁶, R^(11a), R^(11b), R^(12a), R^(12b),R^(13a), R^(13b), R^(14a) and R^(14b) is H.

In one specific embodiment, the compound is of the formula (V), having astructure of formula (Va):

In one embodiment, the invention embraces a compound of the formula (V),where D is a moiety of the formula:

In one variation, the compound is of formula (V), where D is asdescribed in this paragraph and each R⁵, R⁶, R^(11a), R^(11b), R^(12a),R^(12b), R^(13a), R^(13b), R^(14a) and R^(14b) is H. In anothervariation of this embodiment, each R⁵, R⁶, R^(11a), R^(11b), R^(13a),R^(14a) and R^(14b) is H, R^(12a) is CH₃, and R^(12b) and R^(13b) aretaken together to form C(CH₃)₂.

In one embodiment, the invention embraces a compound of the formula(VI):

where D is selected from the group consisting of alkyl-C(O)—,substituted alkyl-C(O)—, perhaloalkyl-C(O)—, alkenyl-C(O)—, substitutedalkenyl-C(O)—, alkynyl-C(O)—, substituted alkynyl-C(O)—, aryl-C(O)—,substituted aryl-C(O)—, heteroaryl-C(O)—, substituted heteroaryl-C(O)—,heterocyclyl-C(O)— and —P(O)(OC₁-C₈alkyl)₂; and

each R^(11a), R^(11b), R^(12a), R^(12b), R^(13a), R^(13b), R^(14a) andR^(14b) is independently H, substituted or unsubstituted C₁-C₈ alkyl,halo, hydroxyl, alkoxy, cyano or nitro, or is taken together with ageminal R group to form a carbonyl moiety, or is taken together withanother R group to form a ring;

or a salt or solvate thereof.

In some embodiments, the compound is of the formula (VI), where at leastone, two, three, four, five, six, seven or eight of R^(11a), R^(11b),R^(12a), R^(12b), R^(13a), R^(13b), R^(14a) and R^(14b) are H. In someembodiments, the compound is of the formula (VI), where one, two, three,four, five, six, seven or eight of R^(11a), R^(11b), R^(12a), R^(12b),R^(13a), R^(13b), R^(14a) and R^(14b) are H. In one specific embodiment,the compound is of the formula (VI), where each R^(11a), R^(11b),R^(12a), R^(12b), R^(13a), R^(13b), R^(14a) and R^(14b) is H.

In one embodiment, the compound is of the formula (VI), where D is amoiety of the formula:

In one variation, each R^(11a), R^(11b), R^(12a), R^(12b), R^(13a),R^(13b), R^(14a) and R^(14b) is H.

In another embodiment, the compound is of the formula (VI), where D is amoiety of the formula:

In one variation of this embodiment, each R^(11a), R^(11b), R^(12a),R^(12b), R^(13a), R^(13b), R^(14a) and R^(14b) is H.

In one variation, the compound is of the formula (VI), having astructure of formula (VIa):

where R³ is unsubstituted or substituted alkyl, perhaloalkyl,unsubstituted or substituted alkenyl, unsubstituted or substitutedalkynyl, unsubstituted or substituted aryl, unsubstituted or substitutedheteroaryl or heterocyclyl.

In one variation, the compound is of the formula (VIa) where R³ is anunsubstituted C₁-C₈ alkyl or a C₁-C₈ alkyl substituted with one or moresubstituents selected from the group consisting of halo, nitro, cyano,oxo, aryl, alkoxy, alkoxycarbonyl, acyl, acyloxy, acylamino, amino,hydroxyl, carboxyl, carboxylalkyl, thiol, thioalkyl, heterocyclyl,—OS(O)₂-alkyl, and the like. In a specific variation, the compound is ofthe formula (VIa) where R³ is a C₁-C₈ alkyl substituted with 1 to 5substituents selected from the group consisting of hydroxyl, alkoxy,acyloxy, acyl, carboxyl, carboxylalkyl, alkoxycarbonyl, andheterocyclyl.

In one specific embodiment, the compound is of the formula (VIb):

where each R^(3a), R^(3b), R^(3c), R^(3d), R^(3e) and R^(3f) isindependently H, halo, hydroxyl, alkoxy, substituted alkoxy, acyl,acyloxy, unsubstituted or substituted alkyl, perhaloalkyl, unsubstitutedor substituted alkenyl, unsubstituted or substituted alkynyl,unsubstituted or substituted aryl, unsubstituted or substitutedheteroaryl, unsubstituted or substituted heterocyclyl, or is takentogether with a geminal R^(3a-f) and the carbon to which they areattached to form a carbonyl; and

R^(3g) is H, unsubstituted or substituted alkyl, perhaloalkyl,unsubstituted or substituted alkenyl, unsubstituted or substitutedalkynyl, unsubstituted or substituted aryl, unsubstituted or substitutedheteroaryl, unsubstituted or substituted heterocyclyl, or is takentogether with one of R^(3e) and R^(3f) to form a lactone moiety;

or a salt or solvate thereof.

In some variations, the compound is of the formula (VIb) where eachR^(3a), R^(3b), R^(3c), R^(3d), R^(3e) and R^(3f) is independently H,unsubstituted or substituted alkyl, hydroxyl, alkoxy, substituted alkoxyor acyloxy. In some variations, the compound is of the formula (VIb)where each R^(3a), R^(3b), R^(3c), R^(3d), R^(3e) and R^(3f) isindependently H, unsubstituted or substituted alkyl, or acyloxy. In somevariations, the compound is of the formula (VIb) where each R^(3a),R^(3b), R^(3c), R^(3d), R^(3e) and R^(3f) is independently H, acyloxy(e.g. acetoxy) or alkyl substituted with an acyloxy group (e.g.acetoxymethyl or nicotinoyloxymethyl). In some variations, at least one,two, three, four, five or six of R^(3a), R^(3b), R^(3c), R^(3d), R^(3e)and R^(3f) is H. In some variations, one, two, three, four, five or sixof R^(3a), R^(3b), R^(3c), R^(3d), R^(3e) and R^(3f) is H. In someparticular variations, one or two of R^(3a), R^(3b), R^(3c), R^(3d),R^(3e) and R^(3f) is independently acyloxy or alkyl substituted with anacyloxy group and each of the remaining R^(3a), R^(3b), R^(3c), R^(3d),R^(3e) and R^(3f) is H. In a particular variation, each R^(3a), R^(3b),R^(3c), R^(3d), R^(3e) and R^(3f) is H.

In some variations, the compound is of the formula (VIb) where R^(3g) isH unsubstituted or substituted alkyl, perhaloalkyl, unsubstituted orsubstituted alkenyl, unsubstituted or substituted alkynyl, unsubstitutedor substituted aryl, unsubstituted or substituted heteroaryl, orunsubstituted or substituted heterocyclyl. In some variations, thecompound is of the formula (VIb) where R^(3g) is unsubstituted orsubstituted alkyl, perhaloalkyl, unsubstituted or substituted aryl,unsubstituted or substituted heteroaryl, or unsubstituted or substitutedheterocyclyl. In some variations, the compound is of the formula (VIb)where R^(3g) is unsubstituted alkyl (e.g. methyl). In some variations,the compound is of the formula (VIb) where R^(3g) is an alkylsubstituted with alkoxy or substituted alkoxy (e.g.2-methoxyethoxymethyl), or an alkyl substituted with aryl or substitutedaryl (e.g. 1-(4-isobutylphenyl)ethyl). In some variations, the compoundis of the formula (VIb) where R^(3g) is a substituted or unsubstitutedheteroaryl (e.g. pyridyl). In some specific variations, the compound isof the formula (VIb) where R^(3g) is unsubstituted alkyl, one or two ofR^(3a), R^(3b), R^(3c), R^(3d), R^(3e) and R^(3f) is acyloxy and each ofthe remaining R^(3a), R^(3b), R^(3c), R^(3d), R^(3e) and R^(3f) is H. Ina particular variation, the compound is of the formula (VIb) whereR^(3g) is unsubstituted alkyl, R^(3c) is acyloxy and each R^(3a),R^(3b), R^(3d), R^(3e) and R^(3f) is H. In another particular variation,the compound is of the formula (VIb) where R^(3g) is unsubstituted alkyland each R^(3a), R^(3b), R^(3c), R^(3d), R^(3e) and R^(3f) is H.

In some variations, the compound is of the formula (VIb) where R^(3g) istaken together with one of R^(3e) and R^(3f) to form a lactone moiety.In one such variation, R^(3g) is taken together with one of R^(3e) andR^(3f) to form a five-membered lactone moiety (e.g.5-oxotetrahydrofuan-2-yl). In another such variation, R^(3g) is takentogether with one of R^(3e) and R^(3f) to form a lactone moiety and theother one of R^(3e) and R^(3f) is H. In a particular variation, thecompound is of the formula (VIb) where R^(3g) is taken together with oneof R^(3e) and R^(3f) to form a lactone moiety and each R^(3a), R^(3b),R^(3c), R^(3d) and the other one of R^(3e) and R^(3f) is H. In a moreparticular variation, the compound is of the formula (VIb) where R^(3g)is taken together with one of R^(3e) and R^(3f) to form a five-memberedlactone moiety and each R^(3a), R^(3b), R^(3c), R^(3d) and the other oneof R^(3e) and R^(3f) is H.

In a particular variation, compounds of the formula (VIc) are provided:

where R^(3a), R^(3b), R^(3c), R^(3d), R^(3e) and R^(3f) are as definedfor formula (VIb) or any variation thereof. In one variation of formula(VIc), R^(3e) and R^(3f) are both H and R^(3a), R^(3b), R^(3c), R^(3d)are as defined for formula (VIb). In a further variation of formula(VIc), R^(3e) and R^(3f) are both H and R^(3a), R^(3b), R^(3c), R^(3d)are independently selected from H or a C₁-C₈ substituted alkyl. In oneaspect of formula (VIc), R^(3e) and R^(3f) are both H, R^(3d) is-alkyl-OMe and R^(3a), R^(3b), R^(3c) are as defined for formula (VIb).In one such variation, R^(3d) is —(CH₂)_(n)—OMe where n is an integerfrom 1 to 5.

In another particular variation, compounds of the formula (VId) areprovided:

where R^(3a), R^(3b), R^(3c), and R^(3d) are as defined for formula(VIb) or any variation thereof.

In one embodiment, the invention embraces a compound of the formula(VII):

where D is selected from the group consisting of alkyl-C(O)—,substituted alkyl-C(O)—, perhaloalkyl-C(O)—, alkenyl-C(O)—, substitutedalkenyl-C(O)—, alkynyl-C(O)—, substituted alkynyl-C(O)—, aryl-C(O)—,substituted aryl-C(O)—, heteroaryl-C(O)—, substituted heteroaryl-C(O)—,heterocyclyl-C(O)— and —P(O)(OC₁-C₈alkyl)₂;

R⁴ is H, substituted or unsubstituted C₁-C₈ alkyl, substituted orunsubstituted C₂-C₈ alkenyl, substituted or unsubstituted C₂-C₈ alkynyl,substituted or unsubstituted acyl, alkoxycarbonyl, substituted orunsubstituted heterocyclyl, substituted or unsubstituted aryl,substituted or unsubstituted aralkyl, or sulfonyl; and

each R^(11a), R^(11b), R^(12a), R^(12b), R^(13a), R^(13b), R^(14a) andR^(14b) is independently H, substituted or unsubstituted C₁-C₈ alkyl,halo, hydroxyl, alkoxy, cyano or nitro, or is taken together with ageminal R group to form a carbonyl moiety, or is taken together withanother R group to form a ring;

provided that the compound is other than8-methyl-3-nitroso-8-azabicyclo[3.2.1]octan-3-yl acetate;

or a salt or solvate thereof.

In one variation, compounds of the formula (VII) are provided where eachR^(11a), R^(11b), R^(12a), R^(12b), R^(13a), R^(13b), R^(14a) andR^(14b) is independently H, substituted or unsubstituted C₁-C₈ alkyl,halo, hydroxyl, alkoxy, cyano or nitro, or is taken together with ageminal R group to form a carbonyl moiety.

In some embodiments, the compound is of the formula (VII), where atleast one, two, three, four, five, six, seven or eight of R^(11a),R^(11b), R^(12a), R^(12b), R^(13a), R^(13b), R^(14a) and R^(14b) are H.In some embodiments, the compound is of the formula (VII), where one,two, three, four, five, six, seven or eight of R^(11a), R^(11b),R^(12a), R^(12b), R^(13a), R^(13b), R^(14a) and R^(14b) are H. In onespecific embodiment, the compound is of the formula (VII), where eachR^(11a), R^(11b), R^(12a), R^(12b), R^(13a), R^(13b), R^(14a) andR^(14b) is H.

In one embodiment, the compound is of the formula (VII), where R⁴ is anunsubstituted C₁-C₈ alkyl or an acyl moiety. In another embodiment, thecompound is of the formula (VII), where R⁴ is an unsubstituted C₁-C₄alkyl, C₁-C₈ alkyl-C(O)— or aryl-C(O)—. In one specific embodiment, thecompound of the formula (VII), where R⁴ is methyl, CH₃C(O)— orphenyl-C(O)—. In a further variation, each of R^(11a), R^(11b), R^(12a),R^(12b), R^(13a), R^(13b), R^(14a) and R^(14b) is H and R⁴ is anunsubstituted C₁-C₈ alkyl such as methyl.

In another embodiment, the compound is of the formula (VII), where D isa moiety of the formula:

In some embodiments, the compound is of the formula (VII), where D is amoiety of the formula:

In one variation the compound is of formula (VII), where D is asdescribed in any variations in this paragraph and each R^(11a), R^(11b),R^(12a), R^(12b), R^(13a), R^(13b), R^(14a) and R^(14b) is H. In anothervariation of this embodiment, R⁴ is an unsubstituted C₁-C₈ alkyl or anacyl moiety. In yet another variation of this embodiment, R⁴ is anunsubstituted C₁-C₄ alkyl, C₁-C₈ alkyl-C(O)— or aryl-C(O)—. In onespecific variation of this embodiment, R⁴ is methyl, CH₃C(O)— orphenyl-C(O)— and each R^(11a), R^(11b), R^(12a), R^(12b), R^(13a),R^(13b), R^(14a) and R^(14b) is H.

In one variation, the compound is of the formula (VII), having astructure of formula (VIIa):

where R³ is unsubstituted or substituted alkyl, perhaloalkyl,unsubstituted or substituted alkenyl, unsubstituted or substitutedalkynyl, unsubstituted or substituted aryl, unsubstituted or substitutedheteroaryl or heterocyclyl; and R⁴ is H, substituted or unsubstitutedC₁-C₈ alkyl, substituted or unsubstituted C₂-C₈ alkenyl, substituted orunsubstituted C₂-C₈ alkynyl, substituted or unsubstituted acyl,alkoxycarbonyl, substituted or unsubstituted heterocyclyl, substitutedor unsubstituted aryl, substituted or unsubstituted aralkyl, orsulfonyl. In a specific such variation, R⁴ is substituted orunsubstituted C₁-C₈ alkyl (e.g. methyl) or substituted or unsubstitutedacyl (e.g. acetyl or benzoyl).

In another variation, the compound is of the formula (VII), having astructure of formula (VIIb):

where R³ is unsubstituted or substituted alkyl, perhaloalkyl,unsubstituted or substituted alkenyl, unsubstituted or substitutedalkynyl, unsubstituted or substituted aryl, unsubstituted or substitutedheteroaryl or heterocyclyl.

In one embodiment, the invention embraces a compound of formula (III),(IV), (V), (Va), (VI) or (VII), where D is selected from the groupconsisting of alkyl-C(O)—, substituted alkyl-C(O)—, perhaloalkyl-C(O)—,alkenyl-C(O)—, substituted alkenyl-C(O)—, alkynyl-C(O)—, substitutedalkynyl-C(O)—, aryl-C(O)—, substituted aryl-C(O)—, heteroaryl-C(O)—,substituted heteroaryl-C(O)—, heterocyclyl-C(O)—. In another embodiment,the invention embraces a compound of formula (III), (IV), (V), (Va),(VI) or (VII), where D is P(O)(OC₁-C₈alkyl)₂. In one embodiment, thecompound is of the formula (III), (IV), (V), (Va), (VI) or (VII), whereD is selected from P(O)(OCH₂CH₃)₂ and P(O)(OCH₂CH₂CH₂CH₃)₂.

In one embodiment, the invention embraces a compound of the formula(III), (IV), (V), (Va), (VI) or (VII), where D is unsubstitutedC₁-C₄alkyl-C(O)—, perhaloalkyl-C(O)—, substituted aryl-C(O)—,substituted aralkyl-C(O)—, or substituted C₁-C₄alkyl-C(O)— where thesubstitution is one or more substituents selected from the groupconsisting of halo, cyano, alkoxy, acyloxy, substituted acyloxy [e.g.CH₃OCH₂CH₂OCH₂C(O)O—], acylamino, substituted acylamino, alkylamino,substituted alkylamino, dialkylamino, N-acyl-substituted alkylamino[e.g. (AcOCH₂)₂CHN(Ac)—], N-alkyl-substituted alkylamino [e.g.(AcOCH₂)₂CHN(Me)-], alkoxycarbonylamino [e.g. t-BuOC(O)NH—], substitutedalkoxycarbonylamino [e.g. PhCH₂OC(O)NH—], alkoxycarbonyl, heterocyclyland substituted heterocyclyl. In one variation, D is a di-substitutedC₁-C₄alkyl-C(O)— where the C₁-C₄alkyl-C(O)— is of the formula

In another embodiment, the invention embraces a compound of formula(III), (IV), (V), (Va), (VI) or (VII), where D is a moiety of theformula:

In another embodiment, the compound is of the formula (III), (IV), (V),(Va), (VI) or (VII), where D is a moiety of the formula:

Compounds of formulae (VI) and (VII) have improved water solubility,chemical stability and ability to release HNO in vivo. Ring systems thatinclude a heteroatom in the cyclic ring system result in improvedaqueous solubility properties. Aqueous solubility of compounds describedherein can be measured using methods known in the art.

For any of the compounds of the invention, such as the compounds offormula (I), (Ia), (II), (IIa), (III), (IV), (V), (Va), (VI) or (VII) orother compounds for use in the methods described herein, recitation ordepiction of the parent compound intends and includes all salts andsolvates thereof, where applicable. As such, all salts, such aspharmaceutically acceptable salts and solvates (e.g. hydrates) of acompound are embraced by the invention and described herein the same asif each and every salts or solvate were specifically and individuallylisted.

Representative compounds detailed herein and for use in the methodsinclude, but are not limited to, the compounds listed in Table 1.

TABLE 1 Representative Compounds According to This Invention CompoundNo. Name (Example) Structure 1 1-Methyl-4-nitrosopiperidin-4-yl acetate(Example 3)

2 4-Nitrosotetrahydro-2H-pyran-4- yl acetate (Example 2)

3 1-Acetyl-4-nitrosopiperidin-4-yl acetate (Example 4)

4 1,3-Diethoxy-2-nitrosopropan-2- yl acetate (Example 5)

5 3-Nitrosotetrahydrothiophen-3-yl acetate (Example 6)

6 1-Benzoyl-4-nitrosopiperidin-4- yl acetate (Example 7)

7 1,2,2,6,6-Pentamethyl-4- nitosopiperidin-4-yl acetate (Example 8)

8 2-Nitrosopropane-1,2,3-triyl triacetate (Example 9)

9 1-Nitrosocyclohexyl 2,4- dichlorobenzoate (Example 11)

10 1-Nitrosocyclohexyl isobutyrate (Example 12)

11 1-Methyl-4-nitrosopiperidin-4-yl isobutyrate (Example 13)

12 1-Methyl-4-nitrosopiperidin-4-yl 2,4-difluorobenzoate (Example 14A)

13 1-Nitrosocyclohexyl 2-chloro- 2,2-difluoroacetate (Example 15)

14 1-Nitrosocyclohexyl 4,4,4- trifluoro-3-methylbutanoate (Example 16)

15 4-Nitrosotetrahydro-2H-pyran-4- yl 2,2,2-trifluoroacetate (Example17)

16 4-Nitrosotetrahydro-2H-pyran-4- yl 3,3,3-trifluoropropanoate (Example18)

17 4-Nitrosotetrahydro-2H-pyran-4- yl 4,4,4-trifluorobutanoate (Example19)

18 1-Nitrosocyclohexyl 2,2,3,3,3- pentafluoropropanoate (Example 20)

19 1-Nitrosocyclohexyl 2- cyanoacetate (Example 21)

20 4-Nitrosotetrahydro-2H-pyran-4- yl 2,2,2-trichloroacetate (Example22)

21 4-Nitrosotetrahydro-2H-pyran-4- yl 2,2,3,3,3- pentafluoropropanoate(Example 23)

22 4-Nitrosotetrahydro-2H-pyran-4- yl 2-chloro-2,2-difluoroacetate(Example 24)

23 (S)-4-Nitrosotetrahydro-2H- pyran-4-yl 2-acetamido-3-phenylpropanoate (Example 25)

24 4-Nitrosotetrahydro-2H-pyran-4- yl pivalate (Example 26)

25 Diethyl 1-nitrosocyclohexyl phosphate (Example 27)

26 Dibutyl 1-nitrosocyclohexyl phosphate (Example 28)

27 Dibutyl 1-methyl-4- nitrosopiperidin-4-yl phosphate (Example 29)

28 1-Methyl-4-nitrosopiperidin-4-yl pivalate (Example 30)

29 1,2,2,6,6-Pentamethyl-4- nitrosopiperidin-4-yl pivalate (Example 31)

30 1-Benzoyl-4-nitrosopiperidin-4- yl 2,2,2-trifluoroacetate (Example32)

31 4-Nitrosotetrahydro-2H-pyran-4- yl 2-benzamidoacetate (Example 33)

32 4-Nitrosotetrahydro-2H-pyran-4- yl 2-acetamidopropanoate (Example 34)

33 4-Nitrosotetrahydro-2H-pyran-4- yl 3-(5-oxotetrahydrofuran-2-yl)propanoate (Example 35)

34 Methyl 4-nitrosotetrahydro-2H- pyran-4-yl succinate (Example 36)

35 2-Methyl-2-((4- nitrosotetrahydro-2H-pyran-4-yloxy)carbonyl)propane-1,3-diyl diacetate (Example 37)

36 4-Nitrosotetrahydro-2H-pyran-4- yl 4-acetoxy-3-(acetoxymethyl)butanoate (Example 38)

37 1-Methyl 4-(4-nitrosotetrahydro- 2H-pyran-4-yl) N-[(benzyloxy)carbonyl]aspartate (Example 39)

38 1-tert-Butyl 4-(4- nitrosotetrahydro-2H-pyran-4-yl)N-(tert-butoxycarobnyl)aspartate (Example 40)

39 8-Methyl-3-nitroso-8- azabicyclo[3.2.1]oct-3-yl acetate (Example 10)

40 2-(Methyl(2-(4- nitrosotetrahydro-2H-pyran-4- yloxy)-2-oxoethyl)amino)propane-1,3-diyl diacetate (Example 49)

41 2-(N-(2-(4-Nitrosotetrahydro- 2H-pyran-4-yloxy)-2-oxoethyl)acetamido)propane-1,3- diyl diacetate (Example 50)

42 2-(Acetoxymethyl)-4-(4- nitrosotetrahydro-2H-pyran-4-yloxy)-4-oxobutyl nicotinate (Example 51)

43 2-(2-(4-Nitrosotetrahydro-2H- pyran-4-yloxy)-2-oxoethyl)propane-1,3-diyl dinicotinate (Example 51)

44 4-Nitrosotetrahydro-2H-pyran-4- yl 4-acetoxybutanoate (Example 41)

45 2-((2-(2- Methoxyethoxy)acetoxy)methyl)- 4-(4-nitrosotetrahydro-2H-pyran-4-yloxy)-4-oxobutyl nicotinate (Example 51)

46 4-Nitrosotetrahydro-2H-pyran-4- yl 4-({2-[4-(2-methylpropyl)phenyl]propanoyl} oxy)butanoate (Example 47)

47 4-Nitrosooxan-4-yl (2R)-2- {[(tert- butoxy)carobnyl]amino}propano ate(Example 48)

48 4-Nitrosotetrahydro-2H-pyran-4- yl 4-(acetyloxy)-3-[(acetyloxy)methyl]but-2-enoate (Example 42)

49 1-Methyl-4-nitrosopiperidin-4-yl 4-(acetyloxy)-3-[(acetyloxy)methyl]butanoate (Example 43)

50 4-Nitrosotetrahydro-2H-pyran-4- yl (2S,3S)-2,3,4-tris(acetyloxy)butanoate (Example 44)

51 4-Nitrosotetrahydro-2H-pyran-4- yl 2-(acetyloxy)benzoate (Example 45)

52 4-[(4-Nitrosotetrahydro-2H- pyran-4-yl)oxy]-4-oxobutyl 2-(acetyloxy)benzoate (Example 46)

53 1-Methyl-4-nitrosopiperidin-4-yl 3,5-difluorobenzoate (Example 14B)

54 4-nitrosooxan-4-yl 4-[(2-{1- methyl-5-[(4- methylphenyl)carbonyl]-1H-pyrrol-2- yl}acetyl)oxy]butanoate (Example 51)

55 4-nitrosooxan-4-yl 2- [(2R,3S,4R,5S)-3,4,5,6-tetrakis(acetyloxy)oxan-2- yl]acetate (Example 51)

56 4-nitrosooxan-4-yl 2-amino-3- hydroxypropanoate (Example 51)

57 4-nitrosooxan-4-yl (2E)- 2,3,4,5,6- pentakis(acetyloxy)hex-2-enoate(Example 51)

58 4-nitrosooxan-4-yl 2,3,4,5,6- pentakis(aceyloxy)hexanoate (Example51)

Compounds for Use in the Methods

The methods described may employ a nitroxyl donor compound describedherein. The methods may employ any compounds detailed herein, such ascompounds described in the Brief Summary of the Invention and elsewhere.For example, the methods described may employ a nitroxyl donor compoundof formula (I). In one variation, the method employs a nitroxyl donor ofthe formula (I), including any one or more of: 1-nitrosocycloheptylacetate, 1-nitrosocycloheptyl benzoate,9-nitrosobicyclo[3.3.1]nonan-9-yl acetate,8-methyl-3-nitroso-8-azabicyclo[3.2.1]octan-3-yl acetate, and compoundswhere: (i) R¹ and R² are taken together to form a substituted 6-memberedcarbocyclic moiety where the substituted 6-membered carbocyclic moietyis a monocyclic or bicyclic ring that is substituted with a moietyselected from the group consisting of alkyl, nitroso, acyl, oxime, andsubstituted alkenyl; or (ii) R¹ and R² are taken together to form anunsubstituted or substituted 5 or 6-membered heterocyclic moiety wherethe 5 or 6-membered heterocyclic moiety is dioxane oracyloxy-substituted tetrahydropyan. In addition, the invention embracespharmaceutical compositions comprising a pharmaceutically acceptablecarrier and any compound detailed herein, such as a nitroxyl donorcompound of formula (I), which may include 1-nitrosocycloheptyl acetate,1-nitrosocycloheptyl benzoate, 9-nitrosobicyclo[3.3.1]nonan-9-ylacetate, 8-methyl-3-nitroso-8-azabicyclo[3.2.1]octan-3-yl acetate, andcompounds where (i) R¹ and R² are taken together to form a substituted6-membered carbocyclic moiety, where the substituted 6-memberedcarbocyclic moiety is a monocyclic or bicyclic ring that is substitutedwith a moiety selected from the group consisting of alkyl, nitroso,acyl, oxime, and substituted alkenyl; or (ii) R¹ and R² are takentogether to form an unsubstituted or substituted 5 or 6-memberedheterocyclic moiety, where the 5 or 6-membered heterocyclic moiety isdioxane or acyloxy-substituted tetrahydropyan.

Methods described for treating ischemia/reperfusion injury or cancer mayemploy a nitroxyl donor compound of formula (I)-(VII), including1-nitrosocycloheptyl acetate, 1-nitrosocycloheptyl benzoate,9-nitrosobicyclo[3.3.1]nonan-9-yl acetate or8-methyl-3-nitroso-8-azabicyclo[3.2.1]octan-3-yl acetate; and compoundswhere (i) R¹ or R² is an unsubstituted C₁-C₈ alkyl, e.g. methyl orpropyl, and D is an NSAID moiety; (ii) R¹ and R² are taken together toform an unsubstituted 6-membered carbocyclic moiety and D isn-alkyl-C(O)—, ClCH₂—C(O)—, CCl₃—C(O)—, CF₃—C(O)—, (CH₃)₃C—C(O)—,alkenyl-C(O)—, substituted alkenyl-C(O)—, unsubstituted aryl-C(O)—,mono-substituted aryl-C(O)— or an NSAID moiety; (iii) R¹ and R² aretaken together to form a substituted 6-membered carbocyclic moiety,where the substituted 6-membered carbocyclic moiety is a monocyclic orbicyclic ring that is substituted with a moiety selected from the groupconsisting of alkyl, nitroso, acyl, oxime, and substituted alkenyl; or(iv) R¹ and R² are taken together to form an unsubstituted orsubstituted 5 or 6-membered heterocyclic moiety, where the 5 or6-membered heterocyclic moiety is dioxane or acyloxy-substitutedtetrahydropyan.

For all compounds disclosed herein, where applicable due to the presenceof a stereocenter, the compound is intended to embrace all possiblestereoisomers of the compound depicted or described. Compositionscomprising a compound with at least one stereocenter are also embracedby the invention, and include racemic mixtures or mixtures containing anenantiomeric excess of one enantiomer or single diastereomers ordiastereomeric mixtures. All such isomeric forms of these compounds areexpressly included herein the same as if each and every isomeric formwere specifically and individually listed. The compounds herein may alsocontain linkages (e.g., carbon-carbon bonds) wherein bond rotation isrestricted about that particular linkage, e.g. restriction resultingfrom the presence of a ring or double bond. Accordingly, all cis/transand E/Z isomers are also expressly included in the present invention.The compounds herein may also be represented in multiple tautomericforms, in such instances, the invention expressly includes alltautomeric forms of the compounds described herein, even though only asingle tautomeric form may be represented.

In one variation, the invention provides for a composition ofsubstantially pure compound. “Substantially pure” intends a preparationof the compound that contains no more than 25% of impurity (e.g. byweight %), which impurity may be another compound altogether or adifferent form of the compound (e.g. a different salt or isomer).Percent purity may be assessed by methods known in the art. In onevariation, a preparation of substantially pure compound is providedwhere the preparation contains no more than 15% of impurity. In anothervariation, a preparation of substantially pure compound is providedwhere the preparation contains no more than 10% impurity. In anothervariation, a preparation of substantially pure compound is providedwhere the preparation contains no more than 5% impurity. In anothervariation, a preparation of substantially pure compound is providedwhere the preparation contains no more than 3% impurity. In anothervariation, a preparation of substantially pure compound is providedwhere the preparation contains no more than 1% impurity.

Preferably, compounds of this invention are provided in purified andisolated forms, for example following column chromatography,high-pressure liquid chromatography, recrystallization, or otherpurification techniques. Where particular stereoisomers of compounds ofthis invention are denoted, such stereoisomers preferably aresubstantially free of other stereoisomers.

General Synthetic Methods

The compounds of this invention may be prepared by a number of processesas generally described below and more specifically in the Exampleshereinafter.

The following abbreviations are used herein: dichloromethane (DCM);dimethylsulfoxide (DMSO).

Unless specifically described, starting materials for the reactions areeither commercially available or may be prepare by known procedures. Forexample, many of the starting materials are available from commercialsuppliers such as Sigma-Aldrich. Others may be prepared by proceduresdescribed in standard reference texts such as March's Advanced OrganicChemistry, (John Wiley and Sons) and Larock's Comprehensive OrganicTransformations (VCH Publishers Inc.).

A method of synthesizing an oxime intermediate used in the synthesis ofcompounds of the invention is shown in General Method 1.

General Method 1

To a solution of hydroxylamine hydrochloride in acetonitrile:water (2:1v/v) at ambient temperature is added sodium acetate and a ketone withstirring. After about 3 hours or at reaction completion which may beassessed by known method, the solvents are removed in vacuo and thereaction quenched with potassium carbonate solution. The organics areextracted into DCM, combined, dried over Na₂SO₄ and concentrated invacuo to afford the oxime without need for further purification.

General Method 1A

To a stirred solution of hydroxylamine hydrochloride (1.1 equiv.) inacetonitrile:water (2:1 v/v) at ambient temperature is added sodiumacetate (1.1 equiv.) and a ketone (1 equiv.). The reaction progress ismonitored by TLC and LC-MS and on completion the solvents are removed invacuo and the reaction quenched with potassium carbonate solution. Theorganics are extracted into DCM, combined, dried over Na₂SO₄ andconcentrated in vacuo to afford the oxime without need for furtherpurification.

A method of synthesizing a compound of the invention from an oximeintermediate is descried in General Methods 2-4.

General Method 2

A solution of an oxime in DCM is added dropwise to a solution of leadtetraacetate in DCM at 0° C. A blue colour gradually appears on additionof the oxime solution. Upon complete addition (about 1 hour) thereaction is allowed to warm to ambient temperature and stirring iscontinued for a further 2-3 hours or until reaction completion. Thereaction is quenched by the addition of water, the phases separated andthe organics dried over Na₂SO₄ and concentrated in vacuo to afford theacetoxy-1-nitroso compound which may be purified, e.g. by columnchromatography on silica gel with pentane/ether (depending on nature ofthe oxime) as the eluent to afford the product as a blue oil.

General Method 2A

To a solution of lead tetraacetate (1 equiv.) in DCM (5 vol) at 0° C. isadded a solution of the oxime (1 equiv.) in DCM (5 vol) dropwise. A bluecolor gradually appears on addition of the oxime solution. Upon completeaddition (about 1 hour) the reaction is allowed to warm to ambienttemperature and stirring continued for a further 2-3 hours or untilreaction completion (monitored by TLC and HPLC). The reaction isquenched by the addition of water, the phases separated and the organicsdried over Na₂SO₄ and concentrated in vacuo to afford theacetoxy-1-nitroso compound which is purified, by silica columnchromatography using appropriate solvent mixtures as the eluent toafford the product.

General Method 3

A solution of an oxime in DCM is added dropwise to a solution of leadtetraacetate and an acid in DCM at 0° C. A blue colour gradually appearson addition of the oxime solution. Upon complete addition (about 1 hour)the reaction is allowed to warm to ambient temperature and stirring iscontinued for a further 2-3 hours or until reaction completion. Thereaction is quenched by the addition of water, the phases separated andthe organics dried over Na₂SO₄ and concentrated in vacuo to afford thecrude nitroso compound, which is purified, e.g. by column chromatographyon silica gel with pentane/ether as the eluent to afford the purifiedcompound as a blue oil.

General Method 3A

To a solution of lead tetraacetate (1 equiv.) and an acid (10 equiv.) inDCM (7 vol) at 0° C. is added a solution of an oxime (1 equiv.) in DCM(3 vol) dropwise. A blue color gradually appears on addition of theoxime solution. Upon complete addition (about 1 hour) the reaction isallowed to warm to ambient temperature and stirring is continued for afurther 2-3 hours or until reaction completion (monitored by TLC andHPLC). The reaction is quenched by the addition of water, the phasesseparated and the organics dried over Na₂SO₄ and concentrated in vacuoto afford the crude acyloxy-1-nitroso compound, which is purified bysilica column chromatography using appropriate solvent mixtures as theeluent to afford the product.

General Method 4

To a solution of bis(acyloxy)iodobenzene (which may be synthesizedaccording to the method described in Org. Lett. 2004, 3613-3615) in DCMcooled to 0° C. is added a solution of an oxime in DCM. After about 2hours or until reaction completion, the reaction mixture is concentratedin vacuo and purified, e.g. by column chromatography on silica gel withDCM as the eluent to afford the purified compound as a blue oil.

General Method 4A

To a solution of bis(acyloxy)iodobenzene (1 equiv.) (synthesizedaccording to the method described in Org. Lett. 2004, 3613-3615) in DCM(10 vol) cooled to 0° C. is added a solution of an oxime (1 equiv.) inDCM (1 vol). The reaction progress is monitored by TLC and LC-MS and oncompletion the reaction mixture is concentrated in vacuo and purified,by silica column chromatography using appropriate solvent mixtures asthe eluent to afford the product.

Methods of Using the Compounds and Compositions

The compounds and compositions herein may be used to treat and/orprevent the onset and/or development of a disease or condition that isresponsive to nitroxyl therapy.

The invention embraces methods of administering to an individual(including an individual identified as in need of such treatment) aneffective amount of a compound to produce a desired effect. Identifyinga subject in need of such treatment can be in the judgment of aphysician, clinical staff, emergency response personnel or other healthcare professional and can be subjective (e.g. opinion) or objective(e.g. measurable by a test or diagnostic method).

One embodiment provides a method of modulating (including increasing) invivo nitroxyl levels in an individual in need thereof, the methodcomprising administering to the individual a compound that donatesnitroxyl under physiological conditions or a pharmaceutically acceptablesalt thereof. An individual is in need of nitroxyl modulation if theyhave or are suspected of having or are at risk of having or developing adisease or condition that is responsive to nitroxyl therapy.

Particular diseases or conditions embraced by the methods of theinvention include cardiovascular diseases such as heart failure orconditions and diseases or conditions that implicate or may implicateischemia/reperfusion injury and cancer, e.g. breast, pancreatic,prostate, and colorectal cancer. These methods are described in moredetail below.

Compositions comprising a nitroxyl-donating compound of the inventionare embraced by the invention. However, the methods described may usemore than one nitroxyl donating compound; for example, the methods mayemploy Angeli's salt and a nitroxyl donor compound of the presentinvention or two or more nitroxyl donor compounds of the presentinvention, which may be administered together or sequentially.

Cardiovascular Diseases

Provided herein are methods of treating cardiovascular diseases such asheart failure by administering an effective amount of at least onenitroxyl donor compound to an individual in need thereof. The methodsprovide for treating a cardiovascular disease, such as heart failure, inan individual in need thereof by administering to the individual acompound according to any formulae detailed herein or a pharmaceuticallyacceptable salt thereof. Also provided are methods of administering atherapeutically effective dose of at least one nitroxyl donatingcompound in combination with at least one other positive inotropic agentto an individual in need thereof. Further provided are methods ofadministering a therapeutically effective amount of at least onenitroxyl donating compound to an individual who is receivingbeta-antagonist therapy and who is experiencing heart failure. Methodsare provided herein for administering compounds of the invention incombination with beta-adrenergic agonists to treat heart failure. Suchagonists include dopamine, dobutamine, and isoproterenol, and analogsand derivatives of such compounds. Also provided are methods ofadministering nitroxyl donors to individuals receiving treatment withbeta-antagonizing agents such as propranolol, metoprolol, bisoprolol,bucindolol, and carvedilol. Further, methods are provided herein fortreating specific classifications of heart failure, such as Class IIIheart failure and acute heart failure.

Also embraced by the invention is a method of treating congestive heartfailure (CHF), including acute congestive heart failure, byadministering an effective amount at least one nitroxyl donatingcompound to an individual in need thereof, which individual may beexperiencing heart failure. Also disclosed is a method of treating CHFby administering an effective amount of at least one nitroxyl donatingcompound in combination with an effective amount of at least one otherpositive inotropic agent to an individual in need thereof, whichindividual may be experiencing heart failure. In one variation, theother positive inotrope is a beta-adrenergic agonist, such asdobutamine. The combined administration of a nitroxyl donor and at leastone other positive inotropic agent comprises administering the nitroxyldonor either sequentially with the other positive inotropic agent forexample, the treatment with one agent first and then the second agent,or administering both agents at substantially the same time, whereinthere is an overlap in performing the administration. With sequentialadministration, an individual is exposed to the agents at differenttimes, so long as some amount of the first agent, which is sufficient tobe therapeutically effective in combination with the second agent,remains in the subject when the other agent is administered. Treatmentwith both agents at the same time can involve administration of theagents in the same dose, such as a physically mixed dose, or in separatedoses administered at the same time.

In particular an embodiment, a nitroxyl donor is administered to anindividual experiencing heart failure that is receiving beta-antagonisttherapy. A beta-antagonist (also known as a beta-blocker) includes anycompound that effectively acts as an antagonist at a subject'sbeta-adrenergic receptors, and provides desired therapeutic orpharmaceutical results, such as diminished vascular tone and/or heartrate. A subject who is receiving beta-antagonist therapy is any subjectto whom a beta-antagonist has been administered, and in whom thebeta-antagonist continues to act as an antagonist at the subject'sbeta-adrenergic receptors. In particular embodiments a determination ofwhether a subject is receiving beta-blocking therapy is made byexamination of the subject's medical history. In other embodiments thesubject is screened for the presence of beta-blocking agents by chemicaltests, such as high-speed liquid chromatography as described in Theviset al., Biomed. Chromatogr., 15:393-402 (2001).

The administration of a nitroxyl donating compound either alone, incombination with a positive inotropic agent, or to a subject receivingbeta-antagonist therapy, is used to treat heart failure of allclassifications. In particular embodiments a nitroxyl donating compoundis used to treat early-stage chronic heart failure, such as Class IIheart failure. In other embodiments a nitroxyl donating compound is usedin combination with a positive inotropic agent, such as isoproterenol totreat Class IV heart failure. In still other embodiments a nitroxyldonating compound is used in combination with another positive inotropicagent, such as isoproterenol to treat acute heart failure. In someembodiments, when a nitroxyl donor is used to treat early stage heartfailure, the dose administered is lower than that used to treat acuteheart failure. In other embodiments the dose is the same as is used totreat acute heart failure.

Also provided are methods of treating cardiovascular diseases orconditions that are responsive to nitroxyl therapy, including coronaryobstructions, coronary artery disease (CAD), angina, heart attack,myocardial infarction, high blood pressure, ischemic cardiomyopathy andinfarction, diastolic heart failure, pulmonary congestion, pulmonaryedema, cardiac fibrosis, valvular heart disease, pericardial disease,circulatory congestive states, peripheral edema, ascites, Chagas'disease, ventricular hypertrophy, heart valve disease, heart failure,including but not limited to congestive heart failure such as acutecongestive heart failure and acute decompensated heart failure. Methodsof treating other cardiovascular diseases or conditions are alsoprovided, such as methods of treating pulmonary hypertension or cardiachypertrophy. The methods employ a nitroxyl donating compound alone or incombination with another positive inotropic agent, which may in oneaspect be another nitroxyl donating compound.

Ischemia/Reperfusion Injury

The invention embraces methods of treating or preventing or protectingagainst ischemia/reperfusion injury. In particular, compounds of theinvention are beneficial for individuals at risk for an ischemic event.Thus, provided herein is a method of preventing or reducing the injuryassociated with ischemia/reperfusion by administering an effectiveamount of at least one nitroxyl donating compound to an individual,preferably prior to the onset of ischemia. The methods provide fortreating ischemia/reperfusion injury in an individual in need thereof byadministering to the individual a compound according to any formulaedetailed herein or a pharmaceutically acceptable salt thereof. Acompound of the invention may be administered to an individual afterischemia but before reperfusion. A compound of the invention may also beadministered after ischemia/reperfusion, but where the administrationprotects against further injury. Also provided is a method in which theindividual is demonstrated to be at risk for an ischemic event. Alsodisclosed is a method of administering a nitroxyl donating compound toan organ that is to be transplanted in an amount effective to reduceischemia/reperfusion injury to the tissues of the organ upon reperfusionin the recipient of the transplanted organ.

Nitroxyl donors of the invention may thus be used in methods ofpreventing or reducing injury associated with futureischemia/reperfusion. For example, administration of a nitroxyl donorprior to the onset of ischemia may reduce tissue necrosis (the size ofinfarct) in at-risk tissues. In live subjects this may be accomplishedby administering an effective amount of a nitroxyl donating compound toan individual prior to the onset of ischemia. In organs to betransplanted this is accomplished by contacting the organ with anitroxyl donor prior to reperfusion of the organ in the transplantrecipient. Compositions comprising more than one nitroxyl-donatingcompound also could be used in the methods described, for example,Angeli's salt and a nitroso derivative of the present invention or twoor more nitroso derivatives of the present invention. Thenitroxyl-donating compound also can be used in combination with otherclasses of therapeutic agents that are designed to minimize ischemicinjury, such as beta blockers, calcium channel blockers, anti-platelettherapy or other interventions for protecting the myocardium inindividuals with coronary artery disease.

One method of administering a nitroxyl donor to live subjects includesadministration of the nitroxyl-donating compound prior to the onset ofischemia. This refers only to the onset of each instance of ischemia andwould not preclude performance of the method with subjects who have hadprior ischemic events, i.e., the method also contemplates administrationof nitroxyl-donating compounds to a subject who has had an ischemicevent in the past.

Individuals can be selected who are at risk of a first or subsequentischemic event. Examples include individuals with knownhypercholesterolemia, EKG changes associated with risk of ischemia,sedentary lifestyle, angiographic evidence of partial coronary arteryobstruction, echocardiographic evidence of myocardial damage, or anyother evidence of a risk for a future or additional ischemic event (forexample a myocardial ischemic event, such as a myocardial infarction(MI), or a neurovascular ischemia such as a cerebrovascular accidentCVA). In particular examples of the methods, individuals are selectedfor treatment who are at risk of future ischemia, but who have nopresent evidence of ischemia (such as electrocardiographic changesassociated with ischemia (for example, peaked or inverted T-waves or STsegment elevations or depression in an appropriate clinical context),elevated CKMB, or clinical evidence of ischemia such as crushingsub-sternal chest pain or arm pain, shortness of breath and/ordiaphoresis). The nitroxyl-donating compound also could be administeredprior to procedures in which myocardial ischemia may occur, for examplean angioplasty or surgery (such as a coronary artery bypass graftsurgery). Also embraced is a method of administering a nitroxyl-donatingcompound to an individual at demonstrated risk for an ischemic event.The selection of an individual with such a status could be performed bya variety of methods, some of which are noted above. For example, anindividual with one of more of an abnormal EKG not associated withactive ischemia, prior history of myocardial infarction, elevated serumcholesterol, etc., would be at risk for an ischemic event. Thus, anat-risk individual could be selected by physical testing or elicitingthe potential subject's medical history to determine whether the subjecthas any indications of risk for an ischemic event. If risk isdemonstrated based on the indications discussed above, or any otherindications that one skilled in the art would appreciate, then theindividual would be considered at demonstrated risk for an ischemicevent.

Ischemia/reperfusion may damage tissues other than those of themyocardium and the invention embraces methods of treating or preventingsuch damage. In one variation, the method finds use in reducing injuryfrom ischemia/reperfusion in the tissue of the brain, liver, gut,kidney, bowel, or in any other tissue. The methods preferably involveadministration of a nitroxyl donor to an individual at risk for suchinjury. Selecting a person at risk for non-myocardial ischemia couldinclude a determination of the indicators used to assess risk formyocardial ischemia. However, other factors may indicate a risk forischemia/reperfusion in other tissues. For example, surgery patientsoften experience surgery related ischemia. Thus, individuals scheduledfor surgery could be considered at risk for an ischemic event. Thefollowing risk factors for stroke (or a subset of these risk factors)would demonstrate a subject's risk for ischemia of brain tissue:hypertension, cigarette smoking, carotid artery stenosis, physicalinactivity, diabetes mellitus, hyperlipidemia, transient ischemicattack, atrial fibrillation, coronary artery disease, congestive heartfailure, past myocardial infarction, left ventricular dysfunction withmural thrombus, and mitral stenosis. Ingall, “Preventing ischemicstroke: current approaches to primary and secondary prevention,”Postgrad. Med., 107(6):34-50 (2000). Further, complications of untreatedinfectious diarrhea in the elderly can include myocardial, renal,cerebrovascular and intestinal ischemia. Slotwiner-Nie & Brandt,“Infectious diarrhea in the elderly,” Gastroenterol. Clin. N. Am.,30(3):625-635 (2001). Alternatively, individuals could be selected basedon risk factors for ischemic bowel, kidney or liver disease. Forexample, treatment would be initiated in elderly subjects at risk ofhypotensive episodes (such as surgical blood loss). Thus, subjectspresenting with such an indication would be considered at risk for anischemic event. Also embraced is a method of administering a nitroxyldonating compound of the invention to an individual who has any one ormore of the conditions listed herein, such as diabetes mellitus orhypertension. Other conditions that may result in ischemia such ascerebral arteriovenous malformation would be considered to demonstraterisk for an ischemic event.

The method of administering nitroxyl to organs to be transplantedincludes administration of nitroxyl prior to removal of the organ fromthe donor, for example through the perfusion cannulas used in the organremoval process. If the organ donor is a live donor, for example akidney donor, the nitroxyl donor can be administered to the organ donoras described above for a subject at risk for an ischemic event. In othercases the nitroxyl donor can be administered by storing the organ in asolution comprising the nitroxyl donor. For example, the nitroxyl donorcan be included in the organ preservation solution, such as Universityof Wisconsin “UW” solution, which is a solution comprising hydroxyethylstarch substantially free of ethylene glycol, ethylene chlorohydrin andacetone (see U.S. Pat. No. 4,798,824).

Cancer

The invention embraces methods of treating cancer by administering aneffective amount of at least one nitroxyl donor compound to anindividual having or who is suspected of having a cancerous disease,e.g. cancer. The invention also provides methods of treating cancer byadministering a therapeutically effective dose of at least one nitroxyldonor compound in combination with at least another anti-cancer agent toan individual having cancer. The methods provide for treating cancer inan individual in need thereof by administering to the individual acompound according to any formulae detailed herein or a pharmaceuticallyacceptable salt thereof.

Cancers that may be treated by the method of this invention include:cancers of the head and neck which include tumors of the head, neck,nasal cavity, paranasal sinuses, nasopharynx, oral cavity, oropharynx,larynx, hypopharynx, salivary glands, and paragangliomas; cancers of theliver and biliary tree, particularly hepatocellular carcinoma;intestinal cancers, particularly colorectal cancer; treat ovariancancer; small cell and non-small cell lung cancer; breast cancersarcomas, such as fibrosarcoma, malignant fibrous histiocytoma,embryonal rhabdomysocarcoma, leiomysosarcoma, neurofibrosarcoma,osteosarcoma, synovial sarcoma, liposarcoma, and alveolar soft partsarcoma; neoplasms of the central nervous systems, particularly braincancer; lymphomas such as Hodgkin's lymphoma, lymphoplasmacytoidlymphoma, follicular lymphoma, mucosa-associated lymphoid tissuelymphoma, mantle cell lymphoma, B-lineage large cell lymphoma, Burkitt'slymphoma, and T-cell anaplastic large cell lymphoma. The method oftreating such diseases comprises administering a therapeuticallyeffective amount of a compound of this to a subject. The method may berepeated as necessary.

Compounds of this invention can be administered in combination withother anti-cancer or cytotoxic agents, including alkylating agents,angiogenesis inhibitors, anti-metabolites, DNA cleavers, DNAcrosslinkers, DNA intercalators, DNA minor groove binders, enediynes,heat shock protein 90 inhibitors, histone deacetylase inhibitors,microtubule stabilizers, nucleoside (purine or pyrimidine) analogs,nuclear export inhibitors, proteasome inhibitors, topoisomerase (I orII) inhibitors, tyrosine kinase inhibitors. Specific anti-cancer orcytotoxic agents include .beta.-lapachone, ansamitocin P3, auristatin,bicalutamide, bleomycin, bleomycin, bortezomib, busulfan, calicheamycin,callistatin A, camptothecin, capecitabine, cisplatin, cryptophycins,daunorubicin, docetaxel, doxorubicin, duocarmycin, dynemycin A,etoposide, floxuridine, floxuridine, fludarabine, fluoruracil,gefitinib, gemcitabine, hydroxyurea, imatinib, interferons,interleukins, irinotecan, methotrexate, mitomycin C, oxaliplatin,paclitaxel, spongistatins, suberoylanilide hydroxamic acid (SAHA),thiotepa, topotecan, trichostatin A, vinblastine, vincristine andvindesine.

Pharmaceutical Composition, Dosage Forms and Treatment Regimens

Also included are pharmaceutically acceptable compositions comprising acompound of the invention or pharmaceutically acceptable salt thereofand any of the methods may employ the compounds of the invention as apharmaceutically acceptable composition. A pharmaceutically acceptablecomposition includes one or more of the compounds of the inventiontogether with a pharmaceutical excipient. The pharmaceuticalcompositions of the invention include those suitable for oral, rectal,nasal, topical (including buccal and sublingual), vaginal or parenteral(including subcutaneous, intramuscular, intravenous, intraperitoneal,intracardiac, intradermal, transdermal and intra-tumoral)administration.

Compounds of this invention may be used in a pharmaceutical formulationcomprising a compound of this invention and an excipient. Excipientsthat may be used include carriers, surface active agents, thickening oremulsifying agents, solid binders, dispersion or suspension aids,solubilizers, colorants, flavoring agents, coatings, disintegratingagents, lubricants, sweeteners, preservatives, isotonic agents, andcombinations thereof. The selection and use of suitable excipients istaught in “Remington: The Science and Practice of Pharmacy”, 21st Ed.(Lippincott Williams & Wilkins 2005), the disclosure of which isincorporated herein by reference.

The compounds or compositions may be prepared as any available dosageform. Unit dosage forms are also intended, which includes discrete unitsof the compound or composition such as capsules, sachets or tablets eachcontaining a predetermined amount of the compound; as a powder orgranules; as a solution or a suspension in an aqueous liquid or anon-aqueous liquid; or as an oil-in-water liquid emulsion or awater-in-oil liquid emulsion, or packed in liposomes and as a bolus orthe like.

A tablet containing the compound or composition may be made bycompression or molding, optionally with one or more accessoryingredients. Compressed tablets may be prepared by compressing in asuitable machine the active ingredient in a free-flowing form such as apowder or granules, optionally mixed with a binder, lubricant, inertdiluent, preservative, surface-active or dispersing agent. Moldedtablets may be made by molding in a suitable machine a mixture of thepowdered compound moistened with an inert liquid diluent. The tabletsoptionally may be coated or scored and may be formulated so as toprovide slow or controlled release of the active ingredient therein.Methods of formulating such slow or controlled release compositions ofpharmaceutically active ingredients, such as those herein and othercompounds known in the art, are known in the art and described inseveral issued US patents, some of which include, but are not limitedto, U.S. Pat. Nos. 4,369,174 and 4,842,866, and references citedtherein. Coatings can be used for delivery of compounds to the intestine(see, e.g. U.S. Pat. Nos. 6,638,534, 5,217,720 and 6,569,457, andreferences cited therein). A skilled artisan will recognize that inaddition to tablets, other dosage forms can be formulated to provideslow or controlled release of the active ingredient. Such dosage formsinclude, but are not limited to, capsules, granulations and gel-caps.

Compositions suitable for topical administration include lozengescomprising the ingredients in a flavored basis, usually sucrose andacacia or tragacanth; and pastilles comprising the active ingredient inan inert basis such as gelatin and glycerin, or sucrose and acacia.

Compositions suitable for parenteral administration include aqueous andnon-aqueous sterile injection solutions which may contain anti-oxidants,buffers, bacteriostats and solutes which render the formulation isotonicwith the blood of the intended recipient; and aqueous and non-aqueoussterile suspensions which may include suspending agents and thickeningagents. The formulations may be presented in unit-dose or multi-dosecontainers, for example, sealed ampules and vials, and may be stored ina freeze dried (lyophilized) condition requiring only the addition ofthe sterile liquid carrier, for example water for injections,immediately prior to use. In one variation, the aqueous composition isacidic, having a pH of about 5.5 to about 7.

Extemporaneous injection solutions and suspensions may be prepared fromsterile powders, granules and tablets.

Administration of the compounds or compositions to an individual mayinvolve systemic exposure or may be local administration, such as when acompound or composition is to be administered at the site of interest.Various techniques can be used for providing the subject compositions atthe site of interest, such as via injection, use of catheters, trocars,projectiles, pluronic gel, stems, sustained drug release polymers orother device which provides for internal access. Where an organ ortissue is accessible because of removal from the patient, such organ ortissue may be bathed in a medium containing the subject compositions,the subject compositions may be painted onto the organ, or may beapplied in any convenient way. The methods of the invention embraceadministration of the compounds to an organ to be donated (such as toprevent ischemia/reperfusion injury). Accordingly, organs that areremoved from one individual for transplant into another individual maybe bathed in a medium containing or otherwise exposed to a compound orcomposition as described herein.

The compounds of the invention, such as those of the formulae herein,may be administered in any suitable dosage amount, which may includedosage levels of about 0.0001 to 4.0 grams once per day (or multipledoses per day in divided doses) for adults. Thus, in certain embodimentsof this invention, a compound herein is administered at a dosage of anydosage range in which the low end of the range is any amount between 0.1mg/day and 400 mg/day and the upper end of the range is any amountbetween 1 mg/day and 4000 mg/day (e.g., 5 mg/day and 100 mg/day, 150mg/day and 500 mg/day). In other embodiments, a compound herein, isadministered at a dosage of any dosage range in which the low end of therange is any amount between 0.1 mg/kg/day and 90 mg/kg/day and the upperend of the range is any amount between 1 mg/kg/day and −32 100 mg/kg/day(e.g., 0.5 mg/kg/day and 2 mg/kg/day, 5 mg/kg/day and 20 mg/kg/day). Thedosing interval can be adjusted according to the needs of theindividual. For longer intervals of administration, extended release ordepot formulations can be used. The dosing can be commensurate withintravenous administration. For instance, the compound can beadministered, such as in a pharmaceutical composition that is amenableto intravenous administration, in an amount of between about 0.01μg/kg/min to about 100 μg/kg/min or between about 0.05 μg/kg/min toabout 95 μg/kg/min or between about 0.1 μg/kg/min to about 90 μg/kg/minor between about 1.0 μg/kg/min to about 80 μg/kg/min or between about10.0 μg/kg/min to about 70 μg/kg/min or between about 20 μg/kg/min toabout 60 μg/kg/min or between about 30 μg/kg/min to about 50 μg/kg/minor between about 0.01 μg/kg/min to about 1.0 μg/kg/min or between about0.01 μg/kg/min to about 10 μg/kg/min or between about 0.1 μg/kg/min toabout 1.0 μg/kg/min or between about 0.1 μg/kg/min to about 10 μg/kg/minor between about 1.0 μg/kg/min to about 5 μg/kg/min or between about 70μg/kg/min to about 100 μg/kg/min or between about 80 μg/kg/min to about90 μg/kg/min. In one variation, the compound is administered to anindividual, such as in a pharmaceutical composition that is amenable tointravenous administration, in an amount of at least about 0.01μg/kg/min or at least about 0.05 μg/kg/min or at least about 0.1μg/kg/min or at least about 0.15 μg/kg/min or at least about 0.25μg/kg/min or at least about 0.5 μg/kg/min or at least about 1.0μg/kg/min or at least about 1.5 μg/kg/min or at least about 5.0μg/kg/min or at least about 10.0 μg/kg/min or at least about 20.0μg/kg/min or at least about 30.0 μg/kg/min or at least about 40.0μg/kg/min or at least about 50.0 μg/kg/min or at least about 60.0μg/kg/min or at least about 70.0 μg/kg/min or at least about 80.0μg/kg/min or at least about 90.0 μg/kg/min or at least about 100.0μg/kg/min or more. In another variation, the compound is administered toan individual, such as in a pharmaceutical composition that is amenableto intravenous administration, in an amount of less than about 100.0μg/kg/min or less than about 90.0 μg/kg/min or less than about 80.0μg/kg/min or less than about 80.0 μg/kg/min or less than about 70.0μg/kg/min or less than about 60.0 μg/kg/min or less than about 50.0μg/kg/min or less than about 40.0 μg/kg/min or less than about 30.0μg/kg/min or less than about 20.0 μg/kg/min or less than about 10.0μg/kg/min or less than about 5.0 μg/kg/min or less than about 2.5μg/kg/min or less than about 1.0 μg/kg/min or less than about 0.5μg/kg/min or less than about 0.05 μg/kg/min or less than about 0.15μg/kg/min or less than about 0.1 μg/kg/min or less than about 0.05μg/kg/min or less than about 0.01 μg/kg/min.

The invention further provides kits comprising one or more compounds asdescribed herein. The kits may employ any of the compounds disclosedherein and instructions for use. The compound may be formulated in anyacceptable form. The kits may be used for any one or more of the usesdescribed herein, and, accordingly, may contain instructions for any oneor more of the stated uses (e.g., treating and/or preventing and/ordelaying the onset and/or the development of a disease or condition thatis responsive to nitroxyl therapy, e.g. heart failure,ischemia/reperfusion injury or cancer).

Kits generally comprise suitable packaging. The kits may comprise one ormore containers comprising any compound described herein. Each component(if there is more than one component) can be packaged in separatecontainers or some components can be combined in one container wherecross-reactivity and shelf life permit.

The kits may optionally include a set of instructions, generally writteninstructions, although electronic storage media (e.g., magnetic disketteor optical disk) containing instructions are also acceptable, relatingto the use of component(s) of the methods of the present invention(e.g., treating, preventing and/or delaying the onset and/or thedevelopment of heart disease or ischemia/reperfusion injury). Theinstructions included with the kit generally include information as tothe components and their administration to an individual.

EXAMPLES

The practice of this invention can be further understood by reference tothe following examples, which are provided by way of illustration andnot of limitation.

All NMR were recorded on one of the following instruments; Bruker AVANCE400 MHz spectrometer, Bruker 250 or Bruker 360 operating at ambientprobe temperature using an internal deuterium lock. Chemical shifts arereported in parts per million (ppm) at lower frequency relative totetramethylsilane (TMS). Standard abbreviations are used throughout (s:singlet; br. s: broad singlet; d: doublet; dd: doublet of doublets; t:triplet; q: quartet; quin: quintet; m: multiplet). Coupling constantsare reported in Hertz (Hz).

Example 1 Synthesis of Oxime Intermediates

The following oximes were prepared according to General Method 1:

Example 1A Tetrahydro-pyran-4-one oxime

To a solution of hydroxylamine hydrochloride (1.53 g, 22 mmol) inacetonitrile:water (10 ml:5 ml) at ambient temperature was added sodiumacetate (1.8 g, 22 mmol) and tetrahydro-pyran-4-one (2.0 g, 20 mmol)with stirring. After 3 hours the solvents were removed in vacuo and thereaction quenched with potassium carbonate solution (10 ml). Theorganics were extracted into DCM (3×50 ml), combined, dried over Na₂SO₄and concentrated in vacuo to afford tetrahydro-pyran-4-one oxime withoutneed for further purification (1.72 g, 75% yield). ¹H NMR (360 MHz,DMSO-d₆) δ δ 10.38 (1H, s), 3.67 (2H, t, 5.7 Hz), 3.60 (2H, t, 5.9 Hz),2.48 (2H, t, 5.9 Hz), 2.23 (2H, t, 5.7 Hz).

Example 1B

1-Methyl-piperidin-4-one oxime was prepared from1-methyl-piperidin-4-one and hydroxylamine hydrochloride usingconditions of General Method 1. ¹H NMR (400 MHz, DMSO-d₆) δ 10.31 (1H,s), 2.45 (2H, t, 6.1 Hz), 2.38 (2H, t, 5.9 Hz), 2.31 (2H, t, J=6.0 Hz),2.20 (2H, t, 6.4 Hz), 2.18 (3H, s).

Example 1C

1-Acetyl-piperidin-4-one oxime was prepared from1-acetyl-piperidin-4-one and hydroxylamine using conditions of GeneralMethod 1. ¹H NMR (400 MHz, DMSO-d₆) δ 10.48 (1H, d, 3.2 Hz), 3.43-3.56(4H, m), 2.48-2.56 (1H, m), 2.42 (1H, t, 6.2 Hz), 2.29-2.34 (1H, m),2.17-2.26 (1H, m), 2.03 (3H, d, 4.2 Hz).

Example 1D

1-Benzoyl-piperidin-4-one oxime was prepared from1-benzoyl-piperidin-4-one and hydroxylamine hydrochloride usingconditions of General Method 1. ¹H NMR (400 MHz, DMSO-d₆) δ 10.53 (1H,br. s.), 7.29-7.54 (5H, m), 3.49-3.81 (4H, m), 2.44-2.66 (2H, m),2.17-2.42 (2H, m).

Example 1E

1,3-Diethoxy-propan-2-one oxime was prepared from1,3-diethoxy-propan-2-one and hydroxylamine hydrochloride usingconditions of General Method 1. ¹H NMR (400 MHz, DMSO-d₆) δ 11.06 (1H,s), 4.18 (2H, s), 3.96 (2H, s), 3.41 (4H, quin, 7.0 Hz), 1.10 (6H, q,7.1 Hz).

Example 1F

1,2,2,6,6-Pentamethyl-piperidin-4-one oxime was prepared from1,2,2,6,6-pentamethyl-piperidin-4-one and hydroxylamine hydrochlorideusing conditions of General Method 1. 1H NMR (250 MHz, CHLOROFORM-d) δ2.52 (2H, s), 2.28 (3H, s), 2.21 (2H, s), 1.12 (6H, s), 1.11 (6H, s).

Example 1G

({8-Methyl-8-azabicyclo[3.2.1]octan-3-ylidene}amino)ol was prepared fromtropinone and hydroxylamine hydrochloride using conditions of GeneralMethod 1. ¹H NMR (250 MHz, CHLOROFORM-d) δ 3.20-3.42 (2H, m), 2.90-3.10(1H, m), 2.59 (1H, dd, 15.0, 3.4 Hz), 2.39 (3H, s), 2.08-2.29 (2H, m),1.91-2.08 (2H, m), 1.41-1.72 (2H, m).

Example 1H

3-(Acetyloxy)-2-(hydroxyimino)propyl acetate was prepared from3-(acetyloxy)-2-oxopropyl acetate (Tetrahedron Lett., 2001; 3331-3334)and hydroxylamine hydrochloride using conditions of General Method 1. 1HNMR (250 MHz, CHLOROFORM-d) δ 8.32 (1H, br. s.), 5.02 (2H, s), 4.75 (2H,s), 2.11 (3H, s), 2.10 (3H, s).

Example 2 Preparation of 4-nitrosotetrahydro-2H-pyran-4-yl acetate

4-Nitrosotetrahydro-2H-pyran-4-yl acetate was prepared according toGeneral Method 2. A solution of tetrahydro-pyran-4-one oxime (3.0 g,26.1 mmol) in DCM (50 ml) was added dropwise to a solution of leadtetraacetate (111.57 g, 26.1 mmol) in DCM (100 ml) at 0° C. A blue colorgradually appears on addition of the oxime solution. Upon completeaddition (1 hour) the reaction was allowed to warm to ambienttemperature and stirring was continued for a further 2-3 hours. Thereaction was quenched by the addition of water, the phases separated andthe organics dried over Na₂SO₄ and concentrated in vacuo to afford4-nitrosotetrahydro-2H-pyran-4-yl acetate which was purified by columnchromatography on silica gel with a gradient of pentane:EtOAc as theeluent to afford the title compound as a blue oil in 56% yield. ¹H NMR(250 MHz, CHLOROFORM-d) δ 1.84 (2H, m), 2.18 (2H, m), 2.25 (3H, s), 3.71(2H, m), 4.03 (2H, m).

Example 3 Preparation of 1-methyl-4-nitrosopiperidin-4-yl acetate

1-Methyl-4-nitrosopiperidin-4-yl acetate was prepared from1-methyl-piperidin-4-one oxime and lead tetraacetate using conditions ofGeneral Method 2. ¹H NMR (400 MHz, chloroform-d) δ 2.86-2.91 (2H, m),2.36 (3H, s), 2.29 (2H, s), 2.27 (2H, t, 2.2 Hz), 2.22 (3H, s),1.85-1.91 (2H, m).

Example 4 Preparation of 1-acetyl-4-nitrosopiperidin-4-yl acetate

1-Acetyl-4-nitrosopiperidin-4-yl acetate was prepared from1-acetyl-piperidin-4-one oxime and lead tetraacetate using conditions ofGeneral Method 2. ¹H NMR (400 MHz, chloroform-d) δ 4.51 (1H, ddd, 9.8,8.3, 4.4 Hz), 3.87 (1H, ddd, 10.0, 8.7, 5.0 Hz), 3.41 (1H, dd, 11.5, 3.2Hz), 3.05 (1H, dd, 13.4, 7.8 Hz), 2.26 (3H, s), 2.15 (3H, s), 2.10-2.18(1H, m), 1.94-2.02 (2H, m), 1.81-1.90 (1H, m).

Example 5 Preparation of 1,3-diethoxy-2-nitrosopropan-2-yl acetate

1,3-Diethoxy-2-nitrosopropan-2-yl acetate was prepared from1,3-diethoxy-propan-2-one oxime and lead tetraacetate using conditionsof General Method 2. ¹H NMR (250 MHz, chloroform-d) δ 4.09 (4H, q),3.16-3.68 (4H, m), 2.17 (3H, s), 1.06 (6H, t).

Example 6 Preparation of 3-nitrosotetrahydrothiophen-3-yl acetate

3-Nitrosotetrahydrothiophen-3-yl acetate was prepared fromtetrahydrothiophen-3-one oxime and lead tetraacetate using conditions ofGeneral Method 2. ¹H NMR (400 MHz, chloroform-d) δ 3.69 (1H, d, 13.2Hz), 3.11 (1H, d, 13.2 Hz), 2.99-3.05 (2H, m), 2.50-2.61 (1H, m),2.22-2.29 (1H, m), 2.22 (3H, s).

Example 7 Preparation of 1-benzoyl-4-nitrosopiperidin-4-yl acetate

1-Benzoyl-4-nitrosopiperidin-4-yl acetate was prepared from1-benzoyl-piperidin-4-one oxime and lead tetraacetate using conditionsof General Method 2. ¹H NMR (400 MHz, chloroform-d) δ 7.42-7.48 (5H, m),4.40-4.86 (1H, m), 3.63-4.13 (1H, m), 3.14-3.52 (2H, m), 2.26 (3H, s),2.05-2.19 (2H, m), 1.80-2.04 (2H, m).

Example 8 Preparation of 1,2,2,6,6-pentamethyl-4-nitrosopiperidin-4-ylacetate

1,2,2,6,6-Pentamethyl-4-nitrosopiperidin-4-yl acetate was prepared from1,2,2,6,6-pentamethyl-piperidin-4-one oxime and lead tetraacetate usingconditions of General Method 2. 1H NMR (500 MHz, CHLOROFORM-d) δ 2.43(3H, s), 2.29-2.40 (2H, m), 2.20 (3H, s), 1.81 (2H, d, 13.9 Hz), 1.28(6H, s), 1.23 (6H, s).

Example 9 Preparation of 2-nitrosopropane-1,2,3-triyl triacetate

2-Nitrosopropane-1,2,3-triyl triacetate was prepared from3-(acetyloxy)-2-(hydroxyimino)propyl acetate and lead tetraacetate usingconditions of General Method 2. 1H NMR (250 MHz, CHLOROFORM-d) δ4.50-4.92 (4H, m), 2.21 (3H, s), 2.06 (6H, s).

Example 10 Preparation of 8-methyl-3-nitroso-8-azabicyclo[3.2.1]oct-3-ylacetate

8-Methyl-3-nitroso-8-azabicyclo[3.2.1]oct-3-yl acetate was prepared from({8-methyl-8-azabicyclo[3.2.1]octan-3-ylidene}amino)ol and leadtetraacetate using conditions of General Method 2. 1H NMR (500 MHz,CHLOROFORM-d) δ 3.49-3.55 (2H, m), 2.63 (2H, d, 15.3 Hz), 2.50 (3H, s),2.21 (3H, s), 2.12-2.19 (2H, m), 2.07 (3H, s), 1.92-2.01 (2H, m),1.79-1.88 (2H, m).

Example 11 Preparation of 1-nitrosocyclohexyl 2,4-dichlorobenzoate

1-Nitrosocyclohexyl 2,4-dichlorobenzoate was prepared according toGeneral Method 3. A solution of cyclohexanone oxime (2.66 g, 23.5 mmol)in DCM (50 ml) was added dropwise to a solution of lead tetra acetate(10.42 g, 23.5 mmol) and 2,4 dichlorobenzoic acid (45.0 g, 235 mmol) inDCM (300 ml) at 0° C. A blue color gradually appears on addition of theoxime solution. Upon complete addition (1 hour) the reaction was allowedto warm to ambient temperature and stirring was continued for a further2-3 hours. The reaction was quenched by the addition of water, thephases separated and the organics dried over Na₂SO₄ and concentrated invacuo to afford 1-nitrosocyclohexyl 2,4-dichlorobenzoate which waspurified by column chromatography on silica gel with hexane:EtOAc as theeluent to afford the title compound as a blue oil in 9% yield. δ_(H)(400 MHz, DMSO-d₆) δ 8.02 (1H, d, 8.4 Hz), 7.85 (1H, d, 2.0 Hz), 7.64(1H, dd, 8.4, 2.0 Hz), 2.03-2.12 (2H, m), 1.38-1.93 (8H, m).

Example 12 Preparation of 1-nitrosocyclohexyl isobutyrate

1-Nitrosocyclohexyl isobutyrate was prepared from cyclohexanone oxime,lead tetraacetate and isobutyric acid using conditions of General Method3. ¹H NMR (400 MHz, chloroform-d) δ 2.68-2.79 (1H, sept, 7.0 Hz),1.71-1.93 (8H, m), 1.48-1.62 (2H, m), 1.29 (3H, s), 1.27 (3H, s).

Example 13 Preparation of 1-methyl-4-nitrosopiperidin-4-yl isobutyrate

1-Methyl-4-nitrosopiperidin-4-yl isobutyrate was prepared from1-methyl-piperidin-4-one oxime, lead tetraacetate and isobutyric acidusing conditions of General Method 3. ¹H NMR (400 MHz, chloroform-d) δ2.82-2.94 (2H, m), 2.65 (1H, sept, 7.0 Hz), 2.36 (3H, s), 2.23-2.35 (4H,m), 1.83-1.92 (2H, m), 1.28 (3H, s), 1.27 (3H, s).

Example 14A Preparation of 1-methyl-4-nitrosopiperidin-4-yl2,4-difluorobenzoate

1-Methyl-4-nitrosopiperidin-4-yl 2,4-difluorobenzoate was prepared from1-methyl-piperidin-4-one oxime, lead tetraacetate and2,4-difluorobenzoic acid using conditions of General Method 3. ¹H NMR(400 MHz, chloroform-d) δ 7.59 (2H, dd, 7.5, 2.3 Hz), 7.09 (1H, tt, 8.6,2.4 Hz), 2.98 (2H, dt, 11.7, 3.6 Hz), 2.32-2.50 (7H, m), 2.04 (2H, dd,14.2, 2.7 Hz).

Example 14B Preparation of 1-methyl-4-nitrosopiperidin-4-yl3,5-difluorobenzoate

1-Methyl-4-nitrosopiperidin-4-yl 3,5-difluorobenzoate is prepared from1-methyl-piperidin-4-one oxime, lead tetraacetate and3,5-difluorobenzoic acid using conditions of General Method 3.

Example 15 Preparation of 1-nitrosocyclohexyl2-chloro-2,2-difluoroacetate

1-Nitrosocyclohexyl 2-chloro-2,2-difluoroacetate was prepared fromcyclohexanone oxime, lead tetraacetate and 2-chloro-2,2-difluoroaceticacid using conditions of General Method 3. ¹H NMR (250 MHz,chloroform-d) δ 2.13-2.42 (3H, m), 1.79-1.99 (4H, m), 1.51-1.74 (3H, m).

Example 16 Preparation of 1-nitrosocyclohexyl4,4,4-trifluoro-3-methylbutanoate

1-Nitrosocyclohexyl 4,4,4-trifluoro-3-methylbutanoate was prepared fromcyclohexanone oxime, lead tetraacetate and 4,4,4-trifluoro-3-methylbutanoic acid using conditions of General Method 3. ¹H NMR (250 MHz,chloroform-d) δ 2.69-2.92 (1H, m), 2.69-2.92 (1H, m), 2.47 (1H, d, 6.5Hz), 1.71-2.05 (4H, m), 1.42-1.63 (2H, m), 1.27 (3H, d, 6.9 Hz),1.10-1.39 (2H, m), 0.87 (1H, d, 7.0 Hz), 0.78-0.99 (1H, m).

Example 17 Preparation of 4-nitrosotetrahydro-2H-pyran-4-yl2,2,2-trifluoroacetate

4-Nitrosotetrahydro-2H-pyran-4-yl 2,2,2-trifluoroacetate was preparedfrom tetrahydro-pyran-4-one oxime, lead tetraacetate and trifluoroaceticacid using conditions of General Method 3. ¹H NMR (250 MHz,chloroform-d) δ 4.18 (2H, m), 3.75 (2H, m), 2.53 (2H, m), 1.92 (2H, m).

Example 18 Preparation of 4-nitrosotetrahydro-2H-pyran-4-yl3,3,3-trifluoropropanoate

4-Nitrosotetrahydro-2H-pyran-4-yl 3,3,3-trifluoropropanoate was preparedfrom tetrahydro-pyran-4-one oxime, lead tetraacetate and3,3,3-trifluoropropionic acid using conditions of General Method 3. ¹HNMR (360 MHz, chloroform-d) δ 3.94-4.17 (2H, m), 3.54-3.79 (2H, m), 3.37(2H, q), 2.23-2.43 (2H, m), 1.65-1.96 (2H, m).

Example 19 Preparation of 4-nitrosotetrahydro-2H-pyran-4-yl4,4,4-trifluorobutanoate

4-Nitrosotetrahydro-2H-pyran-4-yl 4,4,4-trifluorobutanoate was preparedfrom tetrahydro-pyran-4-one oxime, lead tetraacetate and4,4,4-trifluorobutyric acid using conditions of General Method 3. ¹H NMR(250 MHz, chloroform-d) δ 3.91-4.19 (2H, m), 3.48-3.83 (2H, m),2.71-2.94 (2H, m), 2.36-2.67 (2H, m), 2.15-2.34 (2H, m), 1.63-1.96 (2H,m).

Example 20 Preparation of 1-nitrosocyclohexyl2,2,3,3,3-pentafluoropropanoate

1-Nitrosocyclohexyl 2,2,3,3,3-pentafluoropropanoate was prepared fromcyclohexanone oxime, lead tetraacetate and pentafluoropropionic acidusing conditions of General Method 3. ¹H NMR (360 MHz, chloroform-d) δ2.35-2.31 (2H, m), 1.77-2.00 (4H, m), 1.49-1.72 (4H, m).

Example 21 Preparation of 1-nitrosocyclohexyl 2-cyanoacetate

1-Nitrosocyclohexyl 2-cyanoacetate was prepared from cyclohexanoneoxime, lead tetraacetate and cyanoacetic acid using conditions ofGeneral Method 3. ¹H NMR (250 MHz, chloroform-d) δ 3.62 (2H, s),2.01-2.21 (2H, m), 1.70-1.97 (5H, m), 1.41-1.69 (3H, m).

Example 22 Preparation of 4-nitrosotetrahydro-2H-pyran-4-yl2,2,2-trichloroacetate

4-Nitrosotetrahydro-2H-pyran-4-yl 2,2,2-trichloroacetate was preparedfrom tetrahydro-pyran-4-one oxime, lead tetraacetate and trichloroaceticacid using conditions of General Method 3. ¹H NMR (360 MHz,chloroform-d) δ 3.99-4.19 (2H, m), 3.61-3.94 (2H, m), 2.30-2.61 (2H, m),1.58-2.03 (2H, m).

Example 23 Preparation of 4-nitrosotetrahydro-2H-pyran-4-yl2,2,3,3,3-pentafluoropropanoate

4-Nitrosotetrahydro-2H-pyran-4-yl 2,2,3,3,3-pentafluoropropanoate wasprepared from tetrahydro-pyran-4-one oxime, lead tetraacetate andpentafluoropropionic acid using conditions of General Method 3. ¹H NMR(250 MHz, chloroform-d) δ 4.11-4.30 (2H, m), 3.46-3.83 (2H, m),2.45-2.73 (2H, m), 1.68-2.02 (2H, m).

Example 24 Preparation of 4-nitrosotetrahydro-2H-pyran-4-yl2-chloro-2,2-difluoroacetate

4-Nitrosotetrahydro-2H-pyran-4-yl 2-chloro-2,2-difluoroacetate wasprepared from tetrahydro-pyran-4-one oxime, lead tetraacetate andchlorodifluoroacetic acid using conditions of General Method 3. ¹H NMR(360 MHz, chloroform-d) δ 4.06-4.23 (2H, m), 3.52-3.86 (2H, m),2.27-2.70 (2H, m), 1.72-2.07 (2H, m).

Example 25 Preparation of (S)-4-nitrosotetrahydro-2H-pyran-4-yl2-acetamido-3-phenylpropanoate

(S)-4-Nitrosotetrahydro-2H-pyran-4-yl 2-acetamido-3-phenyl propanoatewas prepared from tetrahydro-pyran-4-one oxime, lead tetraacetate and(S)-2-acetylamino-3-phenyl-propionic acid using conditions of GeneralMethod 3. ¹H NMR (400 MHz, chloroform-d) δ 7.09-7.46 (5H, m), 5.92 (1H,d, 7.3 Hz), 4.93-5.12 (1H, m), 3.86-4.10 (2H, m), 3.43-3.68 (2H, m),3.19-3.36 (2H, m), 2.06-2.68 (2H, m), 1.93-2.08 (3H, m), 1.81-1.93 (1H,m), 1.47-1.69 (1H, m).

Example 26 Preparation of 4-nitrosotetrahydro-2H-pyran-4-yl pivalate

4-Nitrosotetrahydro-2H-pyran-4-yl pivalate was prepared fromtetrahydro-pyran-4-one oxime, lead tetraacetate and trimethylacetic acidusing conditions of General Method 3. ¹H NMR (360 MHz, chloroform-d) δ3.91-4.14 (2H, m), 3.57-3.77 (2H, m), 2.10-2.29 (2H, m), 1.75-1.93 (2H,m), 1.34 (9H, s).

Example 27 Preparation of diethyl 1-nitrosocyclohexyl phosphate

Diethyl 1-nitrosocyclohexyl phosphate was prepared from cyclohexanoneoxime, lead tetraacetate and diethyl phosphate using conditions ofGeneral Method 3. ¹H NMR (400 MHz, benzene-d₆) δ 3.90-4.15 (3H, m),1.24-1.78 (10H, m), 0.88-1.10 (7H, m)

Example 28 Preparation of dibutyl 1-nitrosocyclohexyl phosphate

Dibutyl 1-nitrosocyclohexyl phosphate was prepared from cyclohexanoneoxime, lead tetraacetate and dibutyl phosphate using conditions ofGeneral Method 3. ¹H NMR (400 MHz, chloroform-d) δ 4.12-4.21 (1H, m),4.00 (3H, q, 6.7 Hz), 1.61-1.79 (6H, m), 1.36-1.48 (5H, m), 1.25-1.35(4H, m), 0.86-0.97 (9H, m).

Example 29 Preparation of dibutyl 1-methyl-4-nitrosopiperidin-4-ylphosphate

Dibutyl 1-methyl-4-nitrosopiperidin-4-yl phosphate was prepared from1-methyl-piperidin-4-one oxime, lead tetraacetate and dibutyl phosphateusing conditions of General Method 3. ¹H NMR (360 MHz, chloroform-d) δ4.03-4.17 (4H, m), 2.77-2.88 (2H, m), 2.27-2.40 (6H, m), 2.12-2.24 (4H,m), 1.59-1.83 (6H, m), 1.38 (4H, q, 7.2 Hz).

Example 30 Preparation of 1-methyl-4-nitrosopiperidin-4-yl pivalate

1-Methyl-4-nitrosopiperidin-4-yl pivalate was prepared from 1-methylpiperidin-4-one oxime, lead tetraacetate and trimethylacetic acid usingconditions of General Method 3. 1H NMR (400 MHz, CHLOROFORM-d) δ2.86-3.00 (2H, m), 2.37 (3H, s), 2.21-2.35 (4H, m), 1.82-1.94 (2H, m),1.32 (9H, s).

Example 31 Preparation of 1,2,2,6,6-pentamethyl-4-nitrosopiperidin-4-ylpivalate

1,2,2,6,6-Pentamethyl-4-nitrosopiperidin-4-yl pivalate was prepared from1,2,2,6,6-pentamethyl-piperidin-4-one oxime, lead tetraacetate andtrimethylacetic acid using conditions of General Method 3. 1H NMR (250MHz, CHLOROFORM-d) δ 2.43-2.64 (5H, m), 2.09 (3H, s), 1.91 (2H, d, 14.2Hz), 1.39 (6H, d, 5.9 Hz), 1.30 (9H, s), 1.25 (6H, d, 2.6 Hz).

Example 32 Preparation of 1-benzoyl-4-nitrosopiperidin-4-yl2,2,2-trifluoroacetate

1-Benzoyl-4-nitrosopiperidin-4-yl 2,2,2-trifluoroacetate was preparedaccording to General Method 4. To a solution ofbis(trifluoroacetoxy)iodobenzene (990 mg, 2.3 mmol) in DCM (25 ml)cooled to 0° C. was added a solution of 1-benzoyl-piperidin-4-one oxime(500 mg, 2.3 mmol) in DCM (20 ml). After 2 hours the reaction wasconcentrated in vacuo and purified by column chromatography on silicagel with DCM as the eluent to afford the title compound as a blue oil in32% yield. ¹H NMR (400 MHz, chloroform-d) δ 7.43-7.53 (5H, m), 4.90-4.64(1H, m), 4.19-3.89 (1H, m), 3.49-3.28 (2H, m), 2.64-2.34 (2H, m),2.11-1.84 (2H, m).

Example 33 Preparation of 4-nitrosotetrahydro-2H-pyran-4-yl2-benzamidoacetate

4-Nitrosotetrahydro-2H-pyran-4-yl 2-benzamidoacetate was prepared fromtetrahydro-pyran-4-one oxime and bis(benzoylaminoacetoxy)iodobenzene(synthesized from iodobenzene diacetate and benzoylaminoacetic acid)using conditions of General Method 4. ¹H NMR (400 MHz, chloroform-d) δ7.15-7.28 (5H, m), 4.39 (2H, d, 5.4 Hz), 4.03 (2H, m), 3.71 (2H, m),2.18 (2H, m), 1.84 (2H, m).

Example 34 Preparation of 4-nitrosotetrahydro-2H-pyran-4-yl2-acetamidopropanoate

4-Nitrosotetrahydro-2H-pyran-4-yl 2-acetamidopropanoate was preparedfrom tetrahydro-pyran-4-one oxime andbis(2-acetylamino-propionate)iodobenzene (synthesized from iodobenzenediacetate and 2-acetylamino-propionic acid) using conditions of GeneralMethod 4. ¹H NMR (250 MHz, chloroform-d) δ 6.04 (1H, d, 6.9 Hz),4.54-4.85 (1H, m), 3.60-3.91 (4H, m), 2.68 (2H, t, 5.8 Hz), 2.38 (2H, t,5.6 Hz), 2.03 (3H, s), 1.50-1.63 (3H, m).

Example 35 Preparation of 4-nitrosotetrahydro-2H-pyran-4-yl3-(5-oxotetrahydrofuran-2-yl)propanoate

4-Nitrosotetrahydro-2H-pyran-4-yl-3-(5-oxotetrahydrofuran-2-yl)propanoatewas prepared from tetrahydro-pyran-4-one oxime andbis(3-(5-oxo-tetrahydro-furan-2-yl)-propanato) iodobenzene (synthesizedfrom iodobenzene diacetate and 3-(5-oxo-tetrahydro-furan-2-yl)-propionicacid) using conditions of General Method 4. 1H NMR (250 MHz,CHLOROFORM-d) δ 4.46-4.71 (1H, m), 3.92-4.15 (2H, m), 3.62-3.84 (2H, m),1.77-2.80 (12H, m).

Example 36 Preparation of methyl 4-nitrosotetrahydro-2H-pyran-4-ylsuccinate

Methyl 4-nitrosotetrahydro-2H-pyran-4-yl succinate was prepared fromtetrahydro-pyran-4-one oxime and bis(methyl succinate) iodobenzene(synthesized from iodobenzene diacetate and methyl succinic acid) usingconditions of General Method 4. 1H NMR (250 MHz, CHLOROFORM-d) δ3.94-4.12 (2H, m), 3.59-3.80 (5H, m), 2.79-2.94 (2H, m), 2.62-2.75 (2H,m), 2.09-2.30 (2H, m), 1.73-1.90 (2H, m).

Example 37 Preparation of2-methyl-2-((4-nitrosotetrahydro-2H-pyran-4-yloxy)carbonyl)propane-1,3-diyldiacetate

2-Methyl-2-((4-nitrosotetrahydro-2H-pyran-4-yloxy)carbonyl)propane-1,3-diyldiacetate was prepared from tetrahydro-pyran-4-one oxime andbis(3-acetoxy-2-acetoxymethyl-2-methyl-propionate) iodobenzene(synthesized from iodobenzene diacetate and3-acetoxy-2-acetoxymethyl-2-methyl-propionic acid which was in turnsynthesized using the reported method J. Am. Chem. Soc., 118, 1996,6388-6395) using conditions of General Method 4. 1H NMR (250 MHz,CHLOROFORM-d) δ 4.32 (4H, s), 3.98-4.17 (2H, m), 3.54-3.74 (2H, m),2.28-2.46 (2H, m), 2.13 (6H, s), 1.77-1.94 (2H, m), 1.40 (3H, s).

Example 38 Preparation of 4-nitrosotetrahydro-2H-pyran-4-yl4-acetoxy-3-(acetoxymethyl)butanoate

4-Nitrosotetrahydro-2H-pyran-4-yl 4-acetoxy-3-(acetoxymethyl) butanoatewas prepared from tetrahydro-pyran-4-one oxime andbis(4-acetoxy-3-acetoxymethyl-butyrate) iodobenzene using conditions ofGeneral Method 4. (4-acetoxy-3-acetoxymethyl-butyric acid wassynthesized according to the methods in Tetrahedron: Asymmetry, 1997,4079-4088 and Sov. J. Bioorg. Chem. 1977, 323-324), 1H NMR (250 MHz,CHLOROFORM-d) δ 4.83 (2H, m), 4.63 (2H, m), 2.21 (3H, s), 2.06 (6H, s).

Example 39 Preparation of 1-methyl 4-(4-nitrosotetrahydro-2H-pyran-4-yl)N-[(benzyloxy)carbonyl]aspartate

1-Methyl 4-(4-nitrosotetrahydro-2H-pyran-4-yl)N-[(benzyloxy)carbonyl]aspartate was prepared from tetrahydropyran-4-oneoxime and bis((S)-(+)-3-(benzyloxycarbonyl)-5-oxo-4-oxazolidine)iodobenzene using conditions of General Method 4. 1H NMR (500 MHz,CHLOROFORM-d) δ 7.31-7.42 (4H, m), 5.73 (1H, d, 8.1 Hz), 5.16 (2H, s),4.68-4.74 (1H, m), 4.00-4.08 (2H, m), 3.81 (2H, s), 3.63-3.72 (2H, m),3.06-3.26 (2H, m), 2.20-2.30 (2H, m), 1.75-1.86 (2H, m).

Example 40 Preparation of 1-tert-butyl4-(4-nitrosotetrahydro-2H-pyran-4-yl) N-(tert-butoxycarbonyl)aspartate

1-tert-Butyl 4-(4-nitrosotetrahydro-2H-pyran-4-yl)N-(tert-butoxycarbonyl)aspartate was prepared from tetrahydropyran-4-oneoxime and bis(Boc-Asp-O^(t)Bu)-iodobenzene using conditions of GeneralMethod 4. 1H NMR (500 MHz, CHLOROFORM-d) δ 5.42 (1H, d, 7.7 Hz),4.45-4.53 (1H, m), 3.99-4.09 (2H, m), 3.68-3.76 (2H, m), 3.02-3.18 (2H,m), 2.15-2.30 (2H, m), 1.74-1.93 (2H, m), 1.48 (9H, s), 1.46 (9H, s).

Example 41 Preparation of 4-nitrosotetrahydro-2H-pyran-4-yl4-(acetoxy)butanoate

Benzyl-4-bromobutanoate was synthesized according to the methods in J.Med. Chem., 1996, 39, 5176-5182.

Benzyl-4-(acetoxy)butanoate was synthesized from 4-bromobenzylbutyrateand potassium acetate. To a solution of 4-bromobenzylbutyrate (1.0 g,3.89 mmol) in acetonitrile (25 ml) was added potassium acetate (5.94 g,7.78 mmol). The reaction was heated to 90° C. for 18 hours and monitoredby TLC (4:1 heptane:EtOAc) The mixture was allowed to cool to roomtemperature and concentrated in vacuo. The crude product was extractedwith water:EtOAc and the organic phase was separated, dried over Na₂SO₄,filtered and concentrated in vacuo. The title compound was isolated insufficient purity without need for further purification. (0.8 g, 87%yield). ¹H NMR (250 MHz, DMSO-d₆) δ 7.29-7.41 (5H, m), 5.09 (2H, s),4.01 (2H, t, 6.5 Hz), 2.44 (2H, t, 7.3 Hz), 1.97 (3H, s), 1.85 (2H,quin, 6.9 Hz).

4-Acetoxy-butyric acid was synthesized from 4-acetoxybenzylbutyrate. Toa solution of 4-acetoxybenzylbutyrate (1 g, 4.3 mmol) in EtOH (10 ml)was added palladium on charcoal (50 mg, 10% w:w) under an atmosphere ofH₂. After 30 minutes at atmospheric temperature/pressure completesaponification of the benzyl ester was observed. The compound wasisolated by filtration and concentration in vacuo. (0.66 g, 100% yield).¹H NMR (500 MHz, CHLOROFORM-d) δ 4.13 (2H, t, 6.3 Hz), 2.46 (2H, t, 7.3Hz), 2.06 (3H, s), 1.98 (2H, quin, 6.8 Hz).

4-Nitrosotetrahydro-2H-pyran-4-yl 4-acetoxybutanoate was prepared fromtetrahydropyran-4-one oxime and bis(4-acetoxy-butyrate) iodobenzene(synthesized from iodobenzene diacetate and 4-acetoxy-butyric acid) andusing conditions of General Method 4. ¹H NMR (500 MHz, CHLOROFORM-d) δ4.18 (2H, t, 6.3 Hz), 4.05 (2H, dt, 4.2, 11.6 Hz), 3.71 (2H, td, 2.6,11.4 Hz), 2.62 (2H, t, 7.4 Hz), 2.17-2.27 (2H, m), 2.08 (3H, s),2.01-2.08 (2H, m), 1.85 (2H, dd, 2.4, 14.3 Hz).

Example 42 Preparation of 4-nitrosotetrahydro-2H-pyran-4-yl4-(acetyloxy)-3-[(acetyloxy)methyl]but-2-enoate

tert-Butyl 4-(acetyloxy)-3-[(acetyloxy)methyl]but-2-enoate wassynthesized according to the methods in Tetrahedron: Asymmetry, 1997,4079-4088 and Sov. J. Bioorg. Chem. 1977, 323-324.

4-Acetoxy-3-acetoxymethyl-but-2-enoic acid was synthesized fromtert-butyl 4-(acetyloxy)-3-[(acetyloxy)methyl]but-2-enoate. tert-Butyl4-(acetyloxy)-3-[(acetyloxy)methyl]but-2-enoate (1.3 g, 4.8 mmol) wasstirred in a 20% solution of TFA in DCM (20 ml) for 4 hours at ambienttemperature. The title compound was isolated by concentration of thesolution in vacuo as the TFA salt. (1 g, 4.7 mmol). ¹H NMR (500 MHz,CHLOROFORM-d) δ 6.04 (1H, t, 1.6 Hz), 5.27 (2H, s), 4.78 (2H, s), 2.15(3H, s), 2.11 (3H, s).

4-Nitrosotetrahydro-2H-pyran-4-yl4-(acetyloxy)-3-[(acetyloxy)methyl]but-2-enoate was prepared fromtetrahydropyran-4-one oxime and{[4-(acetyloxy)-3-[(acetyloxy)methyl]but-2-enoyl]oxy}(phenyl)-λ³-iodanyl4-(acetyloxy)-3-[(acetyloxy)methyl]but-2-enoate (synthesized fromiodobenzene diacetate and 4-acetoxy-3-acetoxymethyl-but-2-enoic acid)and using conditions of General Method 4. ¹H NMR (500 MHz, CHLOROFORM-d)δ 6.20 (1H, s), 5.21 (2H, s), 4.81 (2H, s), 4.05 (2H, dt, 4.0, 11.6 Hz),3.74 (2H, td, 2.4, 11.4 Hz), 2.19-2.26 (2H, m), 2.18 (3H, s), 2.10 (3H,s), 1.89 (2H, d, 12.3 Hz).

Example 43 Preparation of 1-methyl-4-nitrosopiperidin-4-yl4-(acetyloxy)-3-[(acetyloxy)methyl]butanoate

1-Methyl-4-nitrosopiperidin-4-yl4-(acetyloxy)-3-[(acetyloxy)methyl]butanoate was prepared from1-methyl-piperidin-4-one oxime andbis(4-acetoxy-3-acetoxymethyl-butyrate) iodobenzene using conditions ofGeneral Method 4. ¹H NMR (250 MHz, CHLOROFORM-d) δ 4.13-4.24 (4H, m),2.36-2.63 (14H, m), 2.10 (3H, s), 2.88-2.94 (1H, m), 2.08 (3H, s),1.92-1.96 (1H, m).

Example 44 Preparation of 4-nitrosotetrahydro-2H-pyran-4-yl(2S,3S)-2,3,4-tris(acetyloxy)butanoate

Triacetyl-eryhronic acid was synthesized according to the methoddetailed in J. Am. Chem. Soc., 1939, 61, 1720-1725

4-Nitrosotetrahydro-2H-pyran-4-yl (2S,3S)-2,3,4 tris(acetyloxy)butanoateenoate was prepared from tetrahydropyran-4-one oxime andbis((2S,3S)-2,3,4 tris(acetyloxy)butanoate) iodobenzene (synthesizedfrom iodobenzene diacetate and triacetyl-eryhronic acid) usingconditions of General Method 4. ¹H NMR (500 MHz, CHLOROFORM-d) δ5.54-5.58 (1H, m), 5.31 (1H, d, 4.1 Hz), 4.36-4.47 (2H, m), 3.92-4.07(2H, m), 3.57-3.82 (2H, m), 2.44-2.31 (1H, m), 2.10-2.27 (1H, m), 2.11(3H, s), 2.10 (3H, s), 2.01 (3H, s), 1.84-1.86 (1H, m), 1.70-1.73 (1H,m).

Example 45 Preparation of 4-nitrosotetrahydro-2H-pyran-4-yl2-(acetyloxy)benzoate

4-Nitrosotetrahydro-2H-pyran-4-yl 2-(acetyloxy)benzoate was preparedfrom tetrahydro-pyran-4-one oxime, lead tetraacetate and Aspirin usingconditions of General Method 3. ¹H NMR (500 MHz, CHLOROFORM-d) δ 8.15(1H, dd, 7.8, 1.5 Hz), 7.65 (1H, td, 7.8, 1.5 Hz), 7.40 (1H, t, 7.6 Hz),7.17 (1H, d, 8.1 Hz), 4.08 (2H, dt, 11.6, 4.1 Hz), 3.79 (2H, td, 11.5,2.4 Hz), 2.21-2.29 (5H, m), 1.96-1.99 (2H, m).

Example 46 Preparation of4-[(4-nitrosotetrahydro-2H-pyran-4-yl)oxy]-4-oxobutyl2-(acetyloxy)benzoate

Benzyl 4-{[2-(acetyloxy)phenyl]carbonyloxy}butanoate was synthesizedfrom benzyl-4-bromo)butanoate, 2-(acetyloxy)benzoic acid and potassiumcarbonate. To a solution of benzyl-4-bromobutanoate (0.5 g, 1.9 mmol) inacetonitrile (5 ml) at ambient temperature was added potassium carbonate(0.28 g, 2.0 mmol) and 2-(acetyloxy)benzoic acid (0.35 g, 1.9 mmol). Thereaction was heated to 90° C. for 18 hours and monitored by TLC (4:1heptane:EtOAc) The mixture was allowed to cool to room temperature andconcentrated in vacuo. The crude product was partitioned between waterand EtOAc and the organic phase was separated, dried over Na₂SO₄,filtered and concentrated in vacuo. The title compound was purified bysilica column chromatography eluting with heptane:EtOAc (4:1, v:v) andisolated as a clear, colorless oil (0.556 g, 81% yield). ¹H NMR (250MHz, DMSO-d₆) δ 7.29-7.41 (5H, m), 5.09 (2H, s), 4.01 (2H, t, 6.5 Hz),2.44 (2H, t, 7.3 Hz), 1.97 (3H, s), 1.85 (2H, quin, 6.9 Hz).

4-{[2-(Acetyloxy)phenyl]carbonyloxy}butanoic acid was synthesized frombenzyl 4-{[2-(acetyloxy)phenyl]carbonyloxy}butanoate. To a solution ofbenzyl 4-{[2-(acetyloxy)phenyl]carbonyloxy}butanoate (1.13 g, 3.1 mmol)in EtOH (10 ml) at ambient temperature was added palladium on charcoal(50 mg, 10% w:w) under an atmosphere of H₂. After 3 hours at atmospherictemperature/pressure complete saponification of the benzyl ester wasobserved. The mixture was filtered, and the filtrate was concentrated invacuo to provide the title compound. (0.83 g, 98% yield). ¹H NMR (500MHz, DMSO-d₆) δ ppm 7.95 (1H, dd, 7.9, 1.6 Hz), 7.68 (1H, td, 7.7, 1.6Hz), 7.41 (1H, t, 7.6 Hz), 7.24 (1H, d, 8.1 Hz), 4.23 (2H, t, 6.5 Hz),2.35 (2H, t, 7.2 Hz), 2.27 (3H, s), 1.90 (2H, quin, 6.9 Hz).

4-[(4-Nitrosotetrahydro-2H-pyran-4-yl)oxy]-4-oxobutyl2-(acetyloxy)benzoate was prepared from tetrahydropyran-4-one oxime andbis(4-{[2-(acetyloxy)phenyl]carbonyloxy}butanoate) iodobenzene(synthesized from 4-{[2-(acetyloxy)phenyl]carbonyloxy}butanoic acid andiodobenzene diacetate) and using conditions of General Method 4. ¹H NMR(250 MHz, MeOD) δ ppm 8.00 (1H, dt, 7.9, 2.0 Hz), 7.53-7.70 (1H, m),7.30-7.46 (1H, m), 7.15 (1H, dd, 8.1, 1.2 Hz), 4.32-4.46 (2H, m),3.88-3.97 (2H, m), 3.54-3.76 (2H, m), 2.63-2.77 (2H, m), 2.25-2.36 (4H,m), 1.97-2.21 (4H, m), 1.69-1.88 (1H, m).

Example 47 Preparation of 4-nitrosotetrahydro-2H-pyran-4-yl4-({2-[4-(2-methylpropyl)phenyl]propanoyl}oxy)butanoate

Benzyl 4-({2-[4-(2-methylpropyl)phenyl]propanoyl}oxy)butanoate wassynthesized from benzyl-4-bromo)butanoate,2-[4-(2-methylpropyl)phenyl]propanoic acid and potassium carbonate. To asolution of benzyl-4-bromobutanoate (2.0 g, 7.8 mmol) in acetonitrile(20 ml) at ambient temperature was added potassium carbonate (1.13 g,8.2 mmol) and 2-[4-(2-methylpropyl)phenyl]propanoic acid (1.6 g, 7.8mmol). The reaction was heated to 90° C. for 18 hours and monitored byTLC (4:1 heptane:EtOAc) The mixture was allowed to cool to roomtemperature and concentrated in vacuo. The crude product was partitionedbetween water and EtOAc and the organic phase was separated, dried overNa₂SO₄, filtered and concentrated in vacuo. The title compound waspurified by silica column chromatography eluting with heptane:EtOAc(4:1, v:v) and isolated as a clear, colorless oil (2.5 g, 84% yield). ¹HNMR (250 MHz, CHLOROFORM-d) δ ppm 7.30-7.46 (5H, m), 7.02-7.23 (4H, m),5.11 (2H, s), 3.99-4.21 (2H, m), 3.68 (1H, q, 7.2 Hz), 2.43 (2H, d, 7.2Hz), 2.27-2.39 (2H, m), 1.73-2.04 (3H, m), 1.48 (3H, d, 7.2 Hz) 0.89(6H, d, 6.6 Hz).

4-({2-[4-(2-Methylpropyl)phenyl]propanoyl}oxy)butanoic acid wassynthesized from benzyl4-({2-[4-(2-methylpropyl)phenyl]propanoyl}oxy)butanoate. To a solutionof benzyl 4-({2-[4-(2-methylpropyl)phenyl]propanoyl}oxy)butanoate (2.5g, 6.5 mmol) in EtOH (25 ml) at ambient temperature was added palladiumon charcoal (80 mg, 10% w:w) under an atmosphere of H₂. After 3 hours atatmospheric temperature/pressure complete saponification of the benzylester was observed. The mixture was filtered, and the filtrate wasconcentrated in vacuo to provide the title compound. (1.89 g, 99%yield). ¹H NMR (250 MHz, CHLOROFORM-d) δ 7.02-7.24 (4H, m), 4.12 (2H, t,6.2 Hz), 3.62-3.76 (1H, m), 2.45 (2H, d, 7.2 Hz), 2.26-2.37 (2H, m),1.77-1.99 (3H, m), 1.49 (3H, d, 7.2 Hz) 0.90 (6H, d, 6.9 Hz).

4-Nitrosotetrahydro-2H-pyran-4-yl4-({2-[4-(2-methylpropyl)phenyl]propanoyl}oxy)butanoate was preparedfrom tetrahydropyran-4-one oxime and bis4-({2-[4-(2methylpropyl)phenyl]propanoyl}oxy)butanoate) iodobenzene(synthesized from 4-({2-[4-(2-methylpropyl)phenyl]propanoyl}oxy)butanoicacid and iodobenzene diacetate) and using conditions of General Method4. ¹H NMR (250 MHz, CHLOROFORM-d) δ 7.16-7.25 (2H, m), 7.03-7.14 (2H,m), 4.15 (2H, dt, 12.3, 6.2 Hz), 3.98-4.09 (2H, m), 3.59-3.79 (3H, m),2.40-2.53 (4H, m), 2.27-2.37 (1H, m), 2.11-2.28 (1H, m), 1.74-2.07 (5H,m), 1.50 (3H, dd, 7.2, 4.3 Hz), 0.90 (6H, d, 6.2 Hz).

Example 48 Preparation of 4-Nitrosooxan-4-yl(2R)-2-{[(tert-butoxy)carbonyl]amino}propanoate

4-Nitrosooxan-4-yl (2R)-2-{[(tert-butoxy)carbonyl]amino}propanoate wasprepared from tetrahydropyran-4-one oxime andbis(2R)-2-{[(tert-butoxy)carbonyl]amino}propanoate) iodobenzene(synthesized from (2R)-2-{[(tert-butoxy)carbonyl]amino}propanoic acid)and iodobenzene diacetate) and using conditions of General Method 4. ¹HNMR (250 MHz, DMSO-d₆) δ 7.51 (1H, d, 7.0 Hz), 4.18 (1H, q, 7.2 Hz),3.85-3.98 (2H, m), 3.59 (2H, m), 1.98-2.20 (2H, m), 1.58-1.81 (2H, m),1.32-1.47 (12H, m).

Example 49 Preparation of 4-nitrosooxan-4-yl2-{[1,3-bis(acetyloxy)propan-2-yl](methyl)amino}acetate

3-(Acetyloxy)-2-oxopropyl acetate was prepared according to the methoddetailed in J. Am. Chem. Soc., 118, 1996, 6388-6395)

Tert-butyl 2-{[1,3-bis(acetyloxy)propan-2-yl](methyl)amino}acetate. To asolution of 3-(acetyloxy)-2-oxopropyl acetate (4.54 g, 26 mmol) indichloroethane (75 ml) was added tert-butyl 2-(methylamino)acetate HCl.After stirring for 10 minutes sodium triacetoxyborohydride (8.3 g, 39mmol) was added and stirring was continued for 18 hours and monitored byTLC (1:1 heptane:EtOAc). The reaction was quenched by the addition ofsaturated sodium hydrogen carbonate solution and the crude product wasextracted into dichloromethane. The organic phase was separated, driedover MgSO₄, filtered and concentrated in vacuo. The title compound waspurified by silica column chromatography eluting with 0-20% EtOAc:Heptto yield the title compound (1.32 g, 17% yield). ¹H NMR (250 MHz,CHLOROFORM-d) δ 4.07-4.33 (4H, m), 3.34 (2H, s), 3.23 (1H, t, 5.9 Hz),2.49 (3H, s), 1.99-2.19 (6H, m), 1.33-1.55 (9H, m).

2-{[1,3-bis(acetyloxy)propan-2-yl](methyl)amino}acetic acid. Tert-butyl2-{[1,3-bis(acetyloxy)propan-2-yl](methyl)amino}acetate (1.32 g, 4.3mmol) was stirred in a solution of 20% TFA:dichloromethane (20 ml) for 5hours. Upon completion of reaction (assessed by LC-MS) the reaction wasconcentrated in vacuo and re-dissolved in dichloromethane (10 ml). TheHCl salt was treated with polymer supported-diisopropylamine (3.5 g) for18 hours and the resin removed by filtration. The resin was washed withMeCN (3×10 ml) and the title compound isolated by concentration invacuo. (1.0 g, 93%) ¹H NMR (500 MHz, CHLOROFORM-d) δ 4.18 (4H, d, 6.4Hz), 3.41 (2H, s), 3.17-3.29 (1H, m), 2.50 (3H, s), 2.11 (6H, s).

4-Nitrosooxan-4-yl-2-{[1,3-bis(acetyloxy)propan-2-yl](methyl)amino}acetatecould be synthesized from tetrahydropyran-4-one oxime andbis(2-{[1,3-bis(acetyloxy)propan-2-yl](methyl)amino}acetic acid)iodobenzene (synthesized from iodobenzene diacetate and2-{[1,3-bis(acetyloxy)propan-2-yl](methyl)amino}acetic acid) and usingconditions of General Method 4.

Example 50 Preparation of 4-nitrosooxan-4-yl2-{N-[1,3-bis(acetyloxy)propan-2-yl]acetamido}acetate

3-(Acetyloxy)-2-oxopropyl acetate was prepared according to the methoddetailed in J. Am. Chem. Soc., 118, 1996, 6388-6395)

Tert-butyl 2-{[1,3-bis(acetyloxy)propan-2-yl]amino}acetate. To asolution of 3-(acetyloxy)-2-oxopropyl acetate (3.0 g, 17.24 mmol) ondichloroethane (70 ml) was added tert-butyl 2-aminoacetate (2.35 ml,17.24 mmol). The reaction was stirred for 90 minutes before sodiumtriacetoxyborohydride (5.5 g, 25.86 mmol) was added and stirring wascontinued for 18 hours. The reaction was quenched by the addition ofsaturated sodium hydrogen carbonate solution and the crude product wasextracted into dichloromethane. The organic phase was separated, driedover MgSO₄, filtered and concentrated in vacuo. The title compound waspurified by silica column chromatography eluting with 0-40% EtOAc:Heptto yield the title compound (1.93 g, 38% yield). ¹H NMR (250 MHz,CHLOROFORM-d) δ 3.96-4.26 (4H, m), 3.30-3.45 (2H, m), 3.06 (1H, quin,5.4 Hz), 2.09 (6H, s), 1.47 (9H, s).

Tert-butyl 2-{N-[1,3-bis(acetyloxy)propan-2-yl]acetamido}acetate. Acetylchloride (0.357 ml, 5.0 mmol) was added dropwise to a stirred solutionof tert-butyl 2-{[1,3-bis(acetyloxy)propan-2-yl]amino}acetate (1.22 g,4.2 mmol) and triethylamine (0.696 ml, 5.0 mmol). After 2 hours thereaction was quenched by the addition of water. The organic phase wasseparated and washed with further aliquots of water before being driedover MgSO₄, filtered and concentrated in vacuo. The title compound waspurified by silica column chromatography eluting with 25% EtOAc:Hept toyield the title compound (0.7 g, 50%). ¹H NMR (250 MHz, CHLOROFORM-d) δ4.04-4.44 (5H, m), 3.98 (1H, s), 3.88 (1H, s), 2.08 (3H, s), 2.05 (6H,s), 1.47 (9H, s).

2-{N-[1,3-bis(acetyloxy)propan-2-yl]acetamido}acetic acid. Tert-butyl2-{N-[1,3-bis(acetyloxy)propan-2-yl]acetamido}acetate (1.32 g, 3.9 mmol)was stirred in a solution of 20% TFA:dichloromethane (20 ml) for 4hours. Upon completion of reaction (assessed by LC-MS) the reaction wasconcentrated in vacuo and azeotroped with dichloroethane to remove anyresidual TFA. (0.95 g, 87%). ¹H NMR (500 MHz, CHLOROFORM-d) δ 4.27-4.41(2H, m), 4.12-4.26 (3H, m), 4.07 (1H, s), 3.74 (1H, s), 2.28 (3H, s)2.10 (3H, s) 2.07 (3H, s).

4-nitrosooxan-4-yl 2-{N-[1,3-bis(acetyloxy)propan-2-yl]acetamido}acetatewas synthesized from tetrahydropyran-4-one oxime andbis(2-{N-[1,3-bis(acetyloxy)propan-2-yl]acetamido}acetic acid)iodobenzene and using conditions of General Method 4. ¹H NMR (500 MHz,CHLOROFORM-d) δ 4.29-4.47 (2H, m), 4.10-4.25 (5H, m), 3.65-3.83 (2H, m),2.47-2.58 (1H, m), 2.34-2.45 (1H, m), 2.21-2.31 (2H, m), 2.20 (3H, s),2.13 (6H, s), 2.04-2.09 (2H, m).

Example 51 Preparation of Compounds 54, 55, 56, 57, 58, 43, 42 and 45

Title compounds may be prepared according to General Method 4. The titlecompounds may also be prepared according to literature methods.

Example 52 HNO Production Via N₂O Quantification

HNO production of the compounds may be determined by UV-Visspectroscopy.

Nitrous oxide is produced via the dimerization and dehydration of HNO,and is the most common marker for HNO production (Fukuto, J. M.;Bartberger, M. D.; Dutton, A. S.; Paolocci, N.; Wink, D. A.; Houk, K. N.Chem. Res. Toxicol. 2005, 18, 790-801). HNO, however, can also bepartially quenched by oxygen to yield a product that does not produceN₂O (See, (a) Mincione, F.; Menabuoni, L.; Briganti, F.; Mincione, G.;Scozzafava, A.; Supuran, C. T. J. Enzyme Inhibition 1998, 13, 267-284and (b) Scozzafava, A.; Supuran, C. T. J. Med. Chem. 2000, 43,3677-3687.) Using Angeli's salt (AS) as a benchmark, the relativeamounts of N₂O released from compounds are examined via GC headspaceanalysis. The ability of compounds to donate nitroxyl at pH 7.4 in PBSbuffer at 37° C. is assessed based on the levels of N₂O released.

Example 52A HNO Production Via N₂O Quantification

Compounds were tested in the assay described in Example 52, with thefollowing modification. Test compounds were assessed with and alsowithout the addition of Pig Liver Esterase (PLE) at 37° C. for 90minutes in PBS buffer at pH 7.4. Certain compounds of Table 1 (e.g.,compounds 1, 5, 7, 10, 15, 16, 17, 18, 20, 21, 22, 23, 25, 26, 31, 33,35, 36, 37, 38, 39, 44, 48, 49 and 50) were tested and showed detectablelevels of HNO. Certain compounds of Table 1 exhibited enhanced HNOproduction in the presence of PLE. Compound stability was alsodetermined by assessing the half-life of the compounds in PBS at 37° C.at pH 7.4 with and without the addition of PLE according to methodsknown in the art, e.g., in PCT publication No. PCT/US2007/006710.

Example 53 Use of an In Vitro Model to Determine the Ability ofCompounds of the Invention to Treat, Prevent and/or Delay the Onsetand/or the Development of a Disease or Condition Responsive to NitroxylTherapy

a. Cardiovascular Diseases or Conditions.

In vitro models of cardiovascular disease can also be used to determinethe ability of any of the compounds described herein to treat, preventand/or delay the onset and/or the development of a cardiovasculardisease or condition in an individual. An exemplary in vitro model ofheart disease is described below.

In-vitro models could be utilized to assess vasorelaxation properties ofthe compounds. Isometric tension in isolated rat thoracic aortic ringsegment can be measured as described previously by Crawford, J. H.,Huang, J, Isbell, T. S., Shiva, S., Chacko, B. K., Schechter, A.,Darley-Usmar, V. M., Kerby, J. D., Lang, J. D., Krauss, D., Ho, C.,Gladwin, M. T., Patel, R. P., Blood 2006, 107, 566-575. Upon sacrificeaortic ring segments are excised and cleansed of fat and adheringtissue. Vessels are then cut into individual ring segments (2-3 mm inwidth) and suspended from a force-displacement transducer in a tissuebath. Ring segments are bathed at 37° C. in a bicarbonate-buffered,Krebs-Henseleit (K-H) solution of the following composition (mM): NaCl118; KCl 4.6; NaHCO₃ 27.2; KH₂PO₄ 1.2; MgSO₄ 1.2; CaCl₂ 1.75; Na₂EDTA0.03; and glucose 11.1 and perfused continuously with 21% O₂/5% CO₂/74%N₂. A passive load of 2 g is applied to all ring segments and maintainedat this level throughout the experiments. At the beginning of eachexperiment, indomethacin-treated ring segments are depolarized with KCl(70 mM) to determine the maximal contractile capacity of the vessel.Rings are then washed extensively and allowed to equilibrate. Forsubsequent experiments, vessels are submaximally contracted (50% of KClresponse) with phenylephrine (PE, 3×10⁻⁸-10⁻⁷ M), and L-NMMA, 0.1 mM, isalso added to inhibit eNOS and endogenous NO production. After tensiondevelopment reaches a plateau, nitroxyl donating compounds are addedcumulatively to the vessel bath and effects on tension monitored.

In vitro models can be utilized to determine the effects of nitroxyldonating compounds in changes in developed force and intracellularcalcium in heart muscles. Developed force and intracellular calcium canbe measured in rat trabeculae from normal or diseased (i.e. rats withcongestive heart failure or hypertrophy) as described previously (Gao WD, Atar D, Backx P H, Marban E. Circ Res. 1995; 76:1036-1048). Rats(Sprague-Dawley, 250-300 g) are used in these experiments. The rats areanesthetized with pentobarbital (100 mg/kg) via intra-abdominalinjection, the heart exposed by mid-sternotomy, rapidly excised andplaced in a dissection dish. The aorta is cannulated and the heartperfused retrograde (˜15 mM/min) with dissecting Krebs-Henseleit (H-K)solution equilibrated with 95% O₂ and 5% CO₂. The dissecting K-Hsolution is composed of (mM): NaCl 120, NaHCO₃ 20, KCl5, MgCl₂ 1.2,glucose 10, CaCl₂ 0.5, and 2,3-butanedione monoximine (BDM) 20, pH7.35-7.45 at room temperature (21-22° C.). Trabeculae from the rightventricle of the heart are dissected and mounted between a forcetransducer and a motor arm and superfused with normal K-H solution (KCl,5 mM) at a rate of ˜10 ml/min and stimulated at 0.5 Hz. Dimensions ofthe muscles are measured with a calibration reticule in the ocular ofthe dissection microscope (×40, resolution ˜10 μm).

Force is measured using a force transducer system and is expressed inmillinewtons per square millimeter of cross-sectional area. Sarcomerelength is measured by laser diffraction. Resting sarcomere length is setat 2.20-2.30 μm throughout the experiments.

Intracellular calcium is measured using the free acid form of fura-2 asdescribed in previous studies (Gao et al., 1994; Backx et al., 1995; Gaoet al., 1998). Fura-2 potassium salt is microinjected iontophoreticallyinto one cell and allowed to spread throughout the whole muscle (via gapjunctions). The tip of the electrode (˜0.2 μm in diameter) is filledwith fura-2 salt (1 mM) and the remainder of the electrode was filledwith 150 mM KCl. After a successful impalement into a superficial cellin non-stimulated muscle, a hyperpolarizing current of 5-10 nA is passedcontinuously for ˜15 min. Fura-2 epifluorescence is measured by excitingat 380 and 340 nm. Fluorescent light is collected at 510 nm by aphotomultiplier tube. The output of photomultiplier is collected anddigitized. Ryanodine (1.0 μM) is used to enable steady-state activation.After 15 min of exposure to ryanodine, different levels of tetanizationsare induced briefly (˜4-8 seconds) by stimulating the muscles at 10 Hzat varied extracellular calcium (0.5-20 mM). All experiments areperformed at room temperature (20-22° C.).

b. Diseases or Conditions Implicating Ischemia/Reperfusion.

In vitro models can also be used to determine the ability of any of thecompounds described herein to treat, prevent and/or delay the onsetand/or the development of a disease or condition implicatingischemia/reperfusion injury in an individual.

c. Cancer

Antitumor activities of the compounds described herein can be assessedusing in vitro proliferation assays of tumor cells using well-knownmethods, such as described in Norris A. J. et al. Intl. J. Cancer 2008,122:1905-1910.

Cells of an appropriate cell line, e.g. human breast cancer cell lineMCF-7, are seeded in 96-well tissue culture microtiter plates at ˜4000cells per well for an overnight incubation. Serial 10-fold dilutions oftest compounds are added, and the cells are incubated for 72 h. The cellviability is determined using the CellTiter-Glo™ Luminescent CellViability Assay (Promega; Madison, Wis.). The IC₅₀ is measured as theconcentration of drug required for inhibiting cell growth by 50%.

Example 54 Use of In Vivo and/or Ex Vivo Models to Determine the Abilityof Compounds of the Invention to Treat, Prevent and/or Delay the Onsetand/or the Development of a Disease or Condition Responsive to NitroxylTherapy

a. Cardiovascular Diseases or Conditions.

In vivo models of cardiovascular disease can also be used to determinethe ability of any of the compounds described herein to treat, preventand/or delay the onset and/or the development of a cardiovasculardisease or condition in an individual. An exemplary animal model ofheart disease is described below.

In vivo cardiovascular effects obtained with a nitroxyl donor compoundmay be assessed in a control (normal) dog. The study is conducted inadult (25 kg) mongrel (male) dogs chronically instrumented for conscioushemodynamic analysis and blood sampling, as previously described(Katori, T.; Hoover, D. B.; Ardell, J. L.; Helm, R. H.; Belardi, D. F.;Tocchetti, C. G.; Forfia, P. R.; Kass, D. A.; Paolocci, N. Circ. Res.2005; 96: 234-243.). Micromanometer transducers in the left ventricleprovide pressure, while right atrial and descending aortic cathetersprovide fluid-pressures and sampling conduits. Endocardialsonomicrometers (anteriorposterior, septal-lateral) measure short-axisdimensions, a pneumatic occluder around the inferior vena cavefacilitated pre-load manipulations for pressure-relation analysis.Epicardial pacing leads are placed on the right atrium, and another pairis placed on the right ventricle free wall linked to a permanentpacemaker to induce rapid pacing-cardiac failure. After 10 days ofrecovery, animals are evaluated at baseline sinus rhythm and with atrialpacing (120-160 bpm). Measurements include conscious hemodynamicrecordings for cardiac mechanics.

Compounds of the invention are administrated to a healthy control dog atthe dose of 1-5 μg/kg/min and the resulting cardiovascular data isobtained.

Demonstration that a compound of the invention improves cardiachemodynamics in hearts with congestive failure: After completingprotocols under baseline conditions, congestive heart failure is inducedby tachypacing (210 bpm×3 weeks, 240 bpm×1 week), as previouslydescribed (Katori, T.; et al. Circ. Res. 2005; 96: 234-243.). Briefly,end-diastolic pressure and dP/dt_(max) are measured weekly to monitorfailure progression. When animals demonstrate a rise in EDP more than2×, and dP/dt_(max) of >50% baseline, they are deemed ready forcongestive heart failure studies.

The values for test compounds are obtained after 15 min continuous i.v.infusion (2.5 or 1.25 μg/kg/min) in control and heart failurepreparations, respectively, both in the absence and in the presence ofvolume restoration. For comparison, the same hemodynamic measurementsare obtained with AS in heart failure preparations.

b. Diseases or Conditions Implicating Ischemia/Reperfusion

Ex-vivo models of ischemia/reperfusion can also be used to determine theability of any of the compounds described herein to treat, preventand/or delay the onset and/or the development of a disease or conditionimplicating ischemia/reperfusion injury in an individual. An exemplaryex vivo model of ischemia/reperfusion injury is described below.

Male Wistar rats are housed in identical cages and allowed access to tapwater and a standard rodent diet ad libitum. Each animal is anesthetizedwith 1 g/kg urethane i.p. 10 min after heparin (2,500 U, i.m.)treatment. The chest is opened, and the heart is rapidly excised, placedin ice-cold buffer solution and weighed. Isolated rat hearts areattached to a perfusion apparatus and retrogradely perfused withoxygenated buffer solution at 37° C. The hearts are instrumented aspreviously described in Rastaldo et al., “P-450 metabolite ofarachidonic acid mediates bradykinin-induced negative inotropic effect,”Am. J. Physiol., 280:H2823-H2832 (2001), and Paolocci et al.“cGMP-independent inotropic effects of nitric oxide and peroxynitritedonors: potential role for nitrosylation,” Am. J. Physiol., 279:H1982-H1988 (2000). The flow is maintained constant (approximately 9mL/min/g wet weight) to reach a typical coronary perfusion pressure of85-90 mm Hg. A constant proportion of 10% of the flow rate is applied bymeans of one of two perfusion pumps (Terumo, Tokyo, Japan) using a 50 mLsyringe connected to the aortic cannula. Drug applications are performedby switching from the syringe containing buffer alone to the syringe ofthe other pump containing the drug (nitroxyl donating compound)dissolved in a vehicle at a concentration 10× to the desired finalconcentration in the heart. A small hole in the left ventricular wallallows drainage of the thebesian flow, and a polyvinyl-chloride balloonis placed into the left ventricle and connected to an electromanometerfor recording of left ventricular pressure (LVP). The hearts areelectrically paced at 280-300 bpm and kept in a temperature-controlledchamber (37° C.). Coronary perfusion pressure (CPP) and coronary floware monitored with a second electromanometer and an electromagneticflow-probe, respectively, both placed along the perfusion line. Leftventricular pressure, coronary flow and coronary perfusion pressure arerecorded using a TEAC R-71 recorder, digitized at 1000 Hz and analyzedoff-line with DataQ-Instruments/CODAS software, which allowquantification of the maximum rate of increase of LVP during systole(dP/dt_(max)).

Hearts are perfused with Krebs-Henseleit solution gassed with 95% O₂ and5% CO₂ of the following composition: 17.7 mM sodium bicarbonate, 127 mMNaCl, 5.1 mM KCl, 1.5 mM CaCl₂, 1.26 mM MgCl₂, 11 mM D-glucose,supplemented with 5 μg/mL lidocaine.

Experimental Compounds. The nitroxyl donors are diluted in bufferimmediately prior to use.

Experimental Protocols. Hearts are allowed to stabilize for 30 min, andbaseline parameters are recorded. Typically, coronary flow is adjustedwithin the first 10 min and kept constant from thereon. After 30 minstabilization, hearts are randomly assigned to one of the treatmentgroups, and subjected to 30 min global, no-flow ischemia, followed by 30min of reperfusion (I/R). Pacing of the hearts is stopped at thebeginning of the ischemic period and restarted after the third minute ofreperfusion.

Hearts in a control group are perfused with buffer for an additional 29min after stabilization. Treated hearts are exposed to a nitroxyl donor(e.g., 1 μM final concentration for about 20 min followed by a 10 minbuffer wash-out period).

In all hearts pacing is suspended at the onset of ischemia and restarted3 minutes following reperfusion. As isolated heart preparations maydeteriorate over time (typically after 2-2.5 hrs perfusion), the re-flowduration is limited to 30 min in order to minimize the effects producedby crystalloid perfusion on heart performance, and consistently withother reports.

Assessment of ventricular function. To obtain the maximal developed LVP,the volume of the intra-ventricular balloon is adjusted to anend-diastolic LVP of 10 mm Hg during the stabilization period, asreported in Paolocci, supra, and Hare et al., “Pertussis toxin-sensitiveG proteins influence nitric oxide synthase III activity and proteinlevels in rat hearts,” J. Clin. Invest., 101: 1424-31 (1998). Changes indeveloped LVP, dP/dt_(max) and the end-diastolic value induced by theI/R protocol are continuously monitored. The difference between theend-diastolic LVP (EDLVP) before the end of the ischemic period andduring pre-ischemic conditions is used as an index of the extent ofcontracture development. Maximal recovery of developed LVP anddP/dt_(max) during reperfusion is compared with respective pre-ischemicvalues.

Assessment of myocardial injury. Enzyme release is a measure of severemyocardial injury that has yet to progress to irreversible cell injury.Samples of coronary effluent (2 mL) are withdrawn with a catheterinserted into the right ventricle via the pulmonary artery. Samples aretaken immediately before ischemia and at 3, 6, 10, 20 and 30 min ofreperfusion. LDH release is measured as previously described byBergmeyer & Bernt, “Methods of Enzymatic Analysis,” Verlag Chemie(1974). Data are expressed as cumulative values for the entire reflowperiod.

To corroborate the data relative to myocardial injury, determined by LDHrelease, infarct areas are also assessed in a blinded fashion. At theend of the course (30 min reperfusion), each heart is rapidly removedfrom the perfusion apparatus, and the LV dissected into 2-3 mmcircumferential slices. Following 15 min of incubation at 37° C. in 0.1%solution of nitro blue tetrazolium in phosphate buffer as described inMa et al., “Opposite effects of nitric oxide and nitroxyl onpostischemic myocardial injury,” Proc. Natl. Acad. Sci., 96:14617-14622(1999), unstained necrotic tissue is separated from the stained viabletissue. The areas of viable and necrotic tissue are carefully separateby and independent observer who is not aware of the origin of thehearts. The weight of the necrotic and non-necrotic tissues is thendetermined and the necrotic mass expressed as a percentage of total leftventricular mass.

Data may be subjected to statistical methods such as ANOVA followed bythe Bonferroni correction for post hoc t tests.

c. Cancer

Anticancer activities of compounds described herein can be assessedusing in vivo mouse xenograft models using methods described in NorrisA. J. et al (Intl. J. Cancer 2008, 122, 1905-1910) and Stoyanovsky, D.A. et al (J. Med. Chem. 2004, 47, 210-217).

Mice are inoculated with appropriate tumor cells by subcutaneousinjection into the lower flank. Therapy can be started after 1-3 weekswhen the tumors have reached an average volume of ˜50-60 mm³. Tumordiameters are measured with digital calipers, and the tumor volume iscalculated. The anti-tumor efficacy of test compounds is assessed bycomparison of tumor size in test group to that in the control group.

Example 55 Use of Human Clinical Trials to Determine the Ability toCombination Therapies of the Invention to Treat, Prevent and/or Delaythe Onset and/or the Development of a Disease or Condition Responsive toNitroxyl Therapy

If desired, any of the compounds described herein can also be tested inhumans to determine the ability of the compound to treat, prevent and/ordelay the onset and/or the development of a disease or conditionresponsive to nitroxyl therapy. Standard methods can be used for theseclinical trials. In one exemplary method, subjects with such a diseaseor condition, such as congestive heart failure, are enrolled in atolerability, pharmacokinetics and pharmacodynamics phase I study of atherapy using the compounds of the invention in standard protocols. Thena phase II, double-blind randomized controlled trial is performed todetermine the efficacy of the compounds using standard protocols.

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity ofunderstanding, it is apparent to those skilled in the art that certainminor changes and modifications will be practiced. Therefore, thedescription and examples should not be construed as limiting the scopeof the invention.

All references, publications, patents, and patent applications disclosedherein are hereby incorporated by reference in their entireties.

1. A compound of formula I

or a pharmaceutically acceptable salt or solvate thereof, wherein: eachR¹ and R² is independently a substituted or unsubstituted C₁-C₈ alkyl,substituted or unsubstituted C₂-C₈ alkenyl, substituted or unsubstitutedC₂-C₈ alkynyl, or R¹ and R² are taken together to form an unsubstitutedor substituted 6 or 7-membered carbocyclic moiety or an unsubstituted orsubstituted 5, 6 or 7-membered heterocyclic moiety; D isP(O)(OC₁-C₈alkyl)₂; provided that (i) when R¹ or R² is an unsubstitutedC₁-C₈ alkyl, the R¹ or R² that is an unsubstituted C₁-C₈ alkyl is otherthan methyl or propyl; (ii) when R¹ and R² are taken together to form asubstituted 6-membered carbocyclic moiety, the substituted 6-memberedcarbocyclic moiety is a monocyclic or bicyclic ring that is substitutedwith a moiety other than alkyl, nitroso, acyl, oxime, and substitutedalkenyl; (iii) when R¹ and R² are taken together to form anunsubstituted or substituted 5 or 6-membered heterocyclic moiety, the 5or 6-membered heterocyclic moiety is a monocyclic or bicyclic ring otherthan dioxane or acyloxy-substituted tetrahydropyan; or (iv) when each R₁and R₂ is a substituted C₁-C₈ alkyl, each R₁ and R₂ is other thantrifluoromethyl.
 2. A compound of formula (VIa)

or a pharmaceutically acceptable salt thereof, wherein R³ is a C₁-C₈alkyl substituted with 1 to 5 substituents independently selected fromthe group consisting of hydroxyl, alkoxy, acyloxy, acyl, carboxyl,carboxylalkyl, alkoxycarbonyl, and heterocyclyl.
 3. A compound offormula (VIb):

or a pharmaceutically acceptable salt thereof, wherein: each R^(3a),R^(3b), R^(3c), R^(3d), R^(3e) and R^(3f) is independently H, halo,hydroxyl, alkoxy, substituted alkoxy, acyl, acyloxy, unsubstituted orsubstituted alkyl, perhaloalkyl, unsubstituted or substituted alkenyl,unsubstituted or substituted alkynyl, unsubstituted or substituted aryl,unsubstituted or substituted heteroaryl, unsubstituted or substitutedheterocyclyl, or is taken together with a geminal R^(3a-f) and thecarbon to which they are attached to form a carbonyl; and R^(3g) is H,unsubstituted or substituted alkyl, perhaloalkyl, unsubstituted orsubstituted alkenyl, unsubstituted or substituted alkynyl, unsubstitutedor substituted aryl, unsubstituted or substituted heteroaryl,unsubstituted or substituted heterocyclyl, or is taken together with oneof R^(3e) and R^(3f) to form a lactone moiety.
 4. A pharmaceuticalcomposition comprising (i) a compound according to claim 1, and (ii) apharmaceutically acceptable carrier.
 5. The pharmaceutical compositionof claim 4 wherein the composition is amenable to parenteraladministration.
 6. A pharmaceutical composition comprising (i) acompound according to claim 2, and (ii) a pharmaceutically acceptablecarrier.
 7. The pharmaceutical composition of claim 6 wherein thecomposition is amenable to parenteral administration.
 8. Apharmaceutical composition comprising (i) a compound according to claim3, and (ii) a pharmaceutically acceptable carrier.
 9. The pharmaceuticalcomposition of claim 8 wherein the composition is amenable to parenteraladministration.
 10. A compound selected from:4-Nitrosotetrahydro-2H-pyran-4-yl acetate (Compound 2);4-Nitrosotetrahydro-2H-pyran-4-yl pivalate (Compound 24); Diethyl1-nitrosocyclohexyl phosphate (Compound 25); Dibutyl 1-nitrosocyclohexylphosphate (Compound 26); Dibutyl 1-methyl-4-nitrosopiperidin-4-ylphosphate (Compound 27); 4-Nitrosotetrahydro-2H-pyran-4-yl3-(5-oxotetrahydrofuran-2-yl)propanoate (Compound 33); Methyl4-nitrosotetrahydro-2H-pyran-4-yl succinate (Compound 34);2-Methyl-2-((4-nitrosotetrahydro-2H-pyran-4-yloxy)carbonyl)propane-1,3-diyldiacetate (Compound 35); 4-Nitrosotetrahydro-2H-pyran-4-yl4-acetoxy-3-(acetoxymethyl)butanoate (Compound 36);2-(Acetoxymethyl)-4-(4-nitrosotetrahydro-2H-pyran-4-yloxy)-4-oxobutylnicotinate (Compound 42);2-(2-(4-Nitrosotetrahydro-2H-pyran-4-yloxy)-2-oxoethyl)propane-1,3-diyldinicotinate (Compound 43); 4-Nitrosotetrahydro-2H-pyran-4-yl4-acetoxybutanoate (Compound 44);2-((2-(2-Methoxyethoxy)acetoxy)methyl)-4-(4-nitrosotetrahydro-2H-pyran-4-yloxy)-4-oxobutylnicotinate (Compound 45);4-Nitrosotetrahydro-2H-pyran-4-yl-4-({2-[4-(2-methylpropyl)phenyl]propanoyl}oxy)-butanoate(Compound 46); 4-Nitrosotetrahydro-2H-pyran-4-yl(2S,3S)-2,3,4-tris(acetyloxy)butanoate (Compound 50);4-[(4-Nitrosotetrahydro-2H-pyran-4-yl)oxy]-4-oxobutyl2-(acetyloxy)benzoate (Compound 52); 4-Nitrosooxan-4-yl4-[(2-{1-methyl-5-[(4-methylphenyl)carbonyl]-1H-pyrrol-2-yl}acetyl)oxy]butanoate(Compound 54); and 4-Nitrosooxan-4-yl2,3,4,5,6-pentakis(acetyloxy)hexanoate (Compound 58); andpharmaceutically acceptable salts thereof.
 11. A pharmaceuticalcomposition comprising (i) a compound according to claim 10, and (ii) apharmaceutically acceptable carrier.
 12. The pharmaceutical compositionof claim 11 wherein the composition is amenable to parenteraladministration.
 13. The compound of claim 1, wherein when each R₁ and R₂is a substituted C₁-C₈ alkyl, then each R₁ and R₂ is other than a halosubstituted C₁-C₈ alkyl.
 14. The compound of claim 1, wherein each R¹and R² is independently a unsubstituted C₁-C₈ alkyl, substituted orunsubstituted C₂-C₈ alkenyl, substituted or unsubstituted C₂-C₈ alkynyl,or R¹ and R² are taken together to form an unsubstituted or substituted6 or 7-membered carbocyclic moiety or an unsubstituted or substituted 5,6 or 7-membered heterocyclic moiety.
 15. The compound of claim 1,wherein R¹ and R² are taken together to form an unsubstituted orsubstituted 6 or 7-membered carbocyclic moiety or an unsubstituted orsubstituted 5, 6 or 7-membered heterocyclic moiety.